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Introduction: Community-acquired pneumonia (CAP) is an infectious disease associated with high mortality worldwide. Although Streptococcus pneumoniae remains the most frequent pathogen in CAP, data from recent studies using molecular tests have shown that respiratory viruses play a key role in adults with pneumonia. The impact of difficult-to-treat pathogens on the outcomes of pneumonia is also important even though they represent only a small proportion of overall cases. Despite improvements in the microbiological diagnosis of CAP in recent decades, the identification of the causative pathogen is often delayed because of difficulties in obtaining good-quality sputum samples, issues in transporting samples, and slow laboratory processes. Therefore, the initial treatment of CAP is usually empirical. Point-of-care testing (POCT) was introduced to avoid treatment delays and reduce reliance on empirical antibiotics. Areas covered: This review summarizes the main scientific evidence on the role of POCT in the diagnosis and management of patients with CAP. The authors searched for articles on POCT in pneumonia on PubMed from inception to 20 January 2024. The references in the identified articles were also searched. Expert opinion: POCT involves rapid diagnostic assays that can be performed at the bedside especially in cases of severe CAP and immunocompromised patients. These tests can produce results that could help guide initial therapy and management. [ABSTRACT FROM AUTHOR]

Introduction: One of the biggest obstacles in diagnosing Implant-Associated Infections is the lack of infection criteria and standardized diagnostic methods. These infections present a wide range of symptoms, and their diagnosis can be hampered by the formation of microbial biofilms on the surface of implants. This study aimed to provide insight into the performance of sonication in the diagnosis of infections associated with Cardiac Implantable Electronic Devices, to help define a consensus on the algorithm for the microbial diagnosis of these infections. Methods: We carried out a systematic review with meta-analysis. The PRISMA methodology guidelines were followed, and an advanced search was carried out in PubMed and Web of Science, which enabled 8 articles to be included in the review, in which a meta-analysis was also carried out. QUADAS-2 was used to assess the risk of bias and effect measures were calculated to assess publication bias. Results: The overall sensitivity of the method was 0.823 (95% CI: 0.682-0.910) and the specificity was 0.632 (95% CI: 0.506-0.743). Discussion: These results suggest that sonication may offer advantages in diagnosing these infections. However, it is essential to approach these findings carefully and take into account the recommendations provided in the EHRA 2019 guidelines. This study highlights the importance of more effective diagnostic approaches for implantable medical device-associated infections to improve the quality of treatment and minimize the risks associated with these challenging medical conditions. [ABSTRACT FROM AUTHOR]

As microbiological tests play an important role in our diagnostic algorithms and clinical approach towards patients at-risk for pulmonary aspergillosis, a good knowledge of the diagnostic possibilities and especially their limitations is extremely important. In this review, we aim to reflect critically on the available microbiological diagnostic modalities for diagnosis of pulmonary aspergillosis and formulate some future prospects. Timely start of adequate antifungal treatment leads to a better patient outcome, but overuse of antifungals should be avoided. Current diagnostic possibilities are expanding, and are mainly driven by enzyme immunoassays and lateral flow device tests for the detection of Aspergillus antigens. Most of these tests are directed towards similar antigens, but new antibodies towards different targets are under development. For chronic forms of pulmonary aspergillosis, anti- Aspergillus IgG antibodies and precipitins remain the cornerstone. More studies on the possibilities and limitations of molecular testing including targeting resistance markers are ongoing. Also, metagenomic next-generation sequencing is expanding our future possibilities. It remains important to combine different test results and interpret them in the appropriate clinical context to improve performance. Test performances may differ according to the patient population and test results may be influenced by timing, the tested matrix, and prophylactic and empiric antifungal therapy. Despite the increasing armamentarium, a simple blood or urine test for the diagnosis of aspergillosis in all patient populations at-risk is still lacking. Research on diagnostic tools is broadening from a pathogen focus on biomarkers related to the patient and its immune system. [ABSTRACT FROM AUTHOR]

Abstract The present study aimed to suggest the replacement of animal blood with human blood in culture media, involving alternative methods and ethical considerations, such as animal welfare, in addition to potential laboratory cost reduction. Characteristics of growth and hemolysis development were compared in different culture media, using both sheep blood and human blood. Blood types from the ABO blood group system were tested, and commercially acquired sheep blood agar was used for comparison. Bacteria of the genus Streptococcus spp., Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli were tested. It was observed that growth in media with type A and O positive blood showed closer similarities to those performed in agar with sheep blood. Depending on the bacterial species, the results were either more positive or not, with faster-growing and less demanding bacteria showing better results than, for example, S. pneumoniae, which demonstrated difficulty in the growth process and hemolysis generation in human blood agar. The research suggests that in some situations, sheep blood could be replaced, especially when the goal is growth and isolation, but may not be as suitable when the objective is to analyze hemolysis or when the studied species is demanding.

The purpose of this study was to analyse both negative results of bacteriological cultures and the coexistence of several pathogens at the same intramammary inflammatory focus within bovine udder parenchyma in order to compare the health status of cows’ udders in 1985 and 2021. Mastitis was diagnosed on the basis of anamnesis, clinical inspection of the udder, milk quality, and somatic cell counts (SCC on the Fossomatic 5000-FC). In 1985, joint bacteriological and mycological diagnosis was based on the guidelines established for bovine mastitis by FIL-IDF (1971), while for 2021, it was based on the National Mastitis Council (NMC) laboratory Handbook on bovine mastitis (Hogan et al., 1999). In the first study (1985), mainly the genus Staphylococcus was isolated, but at present other species may be involved in cases of mastitis. In both periods, both subclinical and clinical cases were registered. In addition, the milk yield of cows was observed to increase over time, with concomitant deterioration of mammary gland immunity. The phenomenon of biofilms was documented in the study, which may be explained not only by the low efficacy of antibiotic therapy against udder disease but also by impaired immunity in cows, as the lymphocytes trapped in the biofilm matrix were inactive with respect to both cellular and humoral responses.

Nosocomial pneumonia is a healthcare-associated infection with significant consequences for the patient and the healthcare system. The efficacy of treatment significantly depends on the timeliness and adequacy of the antibiotic therapy regimen. The growth of resistance of gram-negative pathogens of nosocomial pneumonia to antimicrobial agents increases the risk of prescribing inadequate empirical therapy, which worsens the results of patient treatment. Identification of risk factors for infection with multidrug-resistant microorganisms, careful local microbiological monitoring with detection of resistance mechanisms, implementation of antimicrobial therapy control strategy and use of rational combinations of antibacterial drugs are of great importance. In addition, the importance of using new drugs with activity against carbapenem-resistant strains, including ceftazidime/aviabactam, must be understood. This review outlines the current data on the etiology, features of diagnosis and antibacterial therapy of nosocomial pneumonia.

Abstract Background Hospital-acquired and ventilator-associated-pneumonia (HAP/VAP) are one of the most prevalent health-care associated infections in the intensive care unit (ICU). Culture-independent methods were therefore developed to provide faster route to diagnosis and treatment. Among these, metagenomic next-generation sequencing (mNGS) has shown considerable promise. Methods This proof-of-concept study describes the technical feasibility and evaluates the clinical validity of the mNGS for the detection and characterization of the etiologic agents causing hospital-acquired and ventilator-associated pneumonia. We performed a prospective study of all patients with HAP/VAP hospitalized in our intensive care unit for whom a bronchoalveolar lavage (BAL) was performed between July 2017 and November 2018. We compared BAL fluid culture and mNGS results of these patients. Results A total of 32 BAL fluids were fully analyzed. Of these, 22 (69%) were positive by culture and all pathogens identified were also reported by mNGS. Among the culture-positive BAL samples, additional bacterial species were revealed by mNGS for 12 patients, raising the issue of their pathogenic role (colonization versus coinfection). Among BALF with culture-negative test, 5 were positive in mNGS test. Conclusions This study revealed concordant results for pneumonia panel pathogens between mNGS and culture-positive tests and identified additional pathogens potentially implicated in pneumonia without etiologic diagnosis by culture. mNGS has emerged as a promising methodology for infectious disease diagnoses to support conventional methods. Prospective studies with real-time mNGS are warranted to examine the impact on antimicrobial decision-making and clinical outcome.

Copyright of Revista Española de Quimioterapia is the property of Sociedad Espanola de Quimioterapia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Hospital-acquired and ventilator-associated-pneumonia (HAP/VAP) are one of the most prevalent health-care associated infections in the intensive care unit (ICU). Culture-independent methods were therefore developed to provide faster route to diagnosis and treatment. Among these, metagenomic next-generation sequencing (mNGS) has shown considerable promise. Methods: This proof-of-concept study describes the technical feasibility and evaluates the clinical validity of the mNGS for the detection and characterization of the etiologic agents causing hospital-acquired and ventilator-associated pneumonia. We performed a prospective study of all patients with HAP/VAP hospitalized in our intensive care unit for whom a bronchoalveolar lavage (BAL) was performed between July 2017 and November 2018. We compared BAL fluid culture and mNGS results of these patients. Results: A total of 32 BAL fluids were fully analyzed. Of these, 22 (69%) were positive by culture and all pathogens identified were also reported by mNGS. Among the culture-positive BAL samples, additional bacterial species were revealed by mNGS for 12 patients, raising the issue of their pathogenic role (colonization versus coinfection). Among BALF with culture-negative test, 5 were positive in mNGS test. Conclusions: This study revealed concordant results for pneumonia panel pathogens between mNGS and culture-positive tests and identified additional pathogens potentially implicated in pneumonia without etiologic diagnosis by culture. mNGS has emerged as a promising methodology for infectious disease diagnoses to support conventional methods. Prospective studies with real-time mNGS are warranted to examine the impact on antimicrobial decision-making and clinical outcome. [ABSTRACT FROM AUTHOR]

This minireview describes some of the articles published in the last two years related to innovative technologies including CRISPR-Cas, surface-enhanced Raman spectroscopy, microfluidics, flow cytometry, Fourier transform infrared spectroscopy, and artificial intelligence and their application to microbiological diagnosis, molecular typing and antimicrobial susceptibility testing. In addition, some articles related to resistance to new antimicrobials (ceftazidime-avibactam, meropenem-vaborbactam, imipenem-relebactam, and cefiderocol) are also described. [ABSTRACT FROM AUTHOR]

The purpose of this study was to analyse both negative results of bacteriological cultures and the coexistence of several pathogens at the same intramammary inflammatory focus within bovine udder parenchyma in order to compare the health status of cows' udders in 1985 and 2021. Mastitis was diagnosed on the basis of anamnesis, clinical inspection of the udder, milk quality, and somatic cell counts (SCC on the Fossomatic 5000-FC). In 1985, joint bacteriological and mycological diagnosis was based on the guidelines established for bovine mastitis by FIL-IDF (1971), while for 2021, it was based on the National Mastitis Council (NMC) laboratory Handbook on bovine mastitis (Hogan et al., 1999). In the first study (1985), mainly the genus Staphylococcus was isolated, but at present other species may be involved in cases of mastitis. In both periods, both subclinical and clinical cases were registered. In addition, the milk yield of cows was observed to increase over time, with concomitant deterioration of mammary gland immunity. The phenomenon of biofilms was documented in the study, which may be explained not only by the low efficacy of antibiotic therapy against udder disease but also by impaired immunity in cows, as the lymphocytes trapped in the biofilm matrix were inactive with respect to both cellular and humoral responses. [ABSTRACT FROM AUTHOR]

IntroductionOne of the biggest obstacles in diagnosing Implant-Associated Infections is the lack of infection criteria and standardized diagnostic methods. These infections present a wide range of symptoms, and their diagnosis can be hampered by the formation of microbial biofilms on the surface of implants. This study aimed to provide insight into the performance of sonication in the diagnosis of infections associated with Cardiac Implantable Electronic Devices, to help define a consensus on the algorithm for the microbial diagnosis of these infections.MethodsWe carried out a systematic review with meta-analysis. The PRISMA methodology guidelines were followed, and an advanced search was carried out in PubMed and Web of Science, which enabled 8 articles to be included in the review, in which a meta-analysis was also carried out. QUADAS-2 was used to assess the risk of bias and effect measures were calculated to assess publication bias.ResultsThe overall sensitivity of the method was 0.823 (95% CI: 0.682–0.910) and the specificity was 0.632 (95% CI: 0.506–0.743).DiscussionThese results suggest that sonication may offer advantages in diagnosing these infections. However, it is essential to approach these findings carefully and take into account the recommendations provided in the EHRA 2019 guidelines. This study highlights the importance of more effective diagnostic approaches for implantable medical device-associated infections to improve the quality of treatment and minimize the risks associated with these challenging medical conditions.

The research deals with the study of immuno-microbiological parallels of the genital tract in women with acute uncomplicated pyelonephritis with concomitant chronic pelvic inflammatory diseases. In addition to etiotropic antibacterial therapy, correction of the mucosal immunity state of the genital tract is crucial for the treatment of the disease and prevention of its recurrence. The purpose of the work was to investigate the contamination and state of local immunity of the genital tract in women with acute uncomplicated pyelonephritis with different clinical courses. Methods. 246 women of reproductive age suffering from acute uncomplicated pyelonephritis were studied, divided into three variants of the disease clinical course: group 1 -- mild (n=105), group 2--moderate (n=90), and group 3 -- severe (n=51) courses. The scrapings of the mucous membrane of the cervical canal and washings from the vagina were obtained before the start of antimicrobial therapy. Quantitative determination of pathogens was carried out by sowing the biological material on solid nutrient environment. Identification of detached bacteria was carried out according to Bergey's. Mollicutes (Ureaplasma spp., Mycoplasma hominis) were determined by the cultural-fermentative method using test systems and the polymerase chain reaction. Vaginal washings of 121 sick women were used for immunological studies. The levels of myeloperoxidase, lysozyme, human β-defensin-2, immunoglobulins M, A, G, secretory IgA, lactoferrin, C3-component of complement, secretory component, and tumor necrosis factor-α were determined. The reference group consisted of 23 clinically healthy women. The Statistica 12.0 program package for Windows was used, and a difference of p<0.05 was considered verifiable. Results. In the majority of patients with acute uncomplicated pyelonephritis (85.6% of cases), concomitant chronic pelvic inflammatory diseases (colpitis, salpingitis, underlying medical condition of cervix) were observed. Classical bacteria were more often detected in vaginal washings of patients of all groups, while mollicutes were detected in scrapings of the mucous membrane of the cervical canal, whereas the frequency of their detection increased with increase in the pyelonephritis severity. The analysis of indicators of local immunity determined disorders of mucosal immunity of the genital tract in the form of increased levels of myeloperoxidase, IgA, and IgG relative to reference values. The highest levels of lysozyme and myeloperoxidase were determined in patients with a severe course of acute uncomplicated pyelonephritis (by 5.3 and 3.6 times more, respectively), and the content of immunoglobulins A, M, and G in patients with a mild course of the disease (by 3.2, 3.1, and 4.0 times more, respectively). An increase in the level of tumor necrosis factor-α was detected in all patients, although no significant differences from controls were found in any group, but the highest median value was recorded in patients with severe pyelonephritis. In the group of patients with a severe clinical course, the lowest median was observed for β-defensin-2, which makes it impossible to inhibit the synthesis of TNF-α and, thus, supports the inflammatory process. Conclusions. It has been proven that in women with acute uncomplicated pyelonephritis, the severity of the course probably correlates with infection of the genital tract by mollicutes, mainly in association with classical bacteria and with level of mucosal immunity disorder. The obtained results of microbiological and immunological studies of biological material collected before the start of antibacterial therapy in patients with acute uncomplicated pyelonephritis proved the need for a mandatory examination by a gynecologist to ascertain concomitant chronic pelvic inflammatory diseases and establish infection with mollicutes to provide relevant etiotropic treatment. The above is the basis for adding the data regarding laboratory examination and treatment to the management protocols of patients with acute uncomplicated pyelonephritis. [ABSTRACT FROM AUTHOR]

Resumen: El uso de dispositivos de estimulación cardiaca se ha incrementado de forma significativa en los últimos años, principalmente en pacientes de edad avanzada y con comorbilidades importantes, lo que hace que exista un mayor riesgo de complicaciones, incluida la infección. En este sentido, la detección e identificación del microorganismo causante de la infección es crucial para una correcta elección y duración del tratamiento antimicrobiano. En esta revisión, presentamos la evidencia clínica más reciente sobre la etiología y diagnóstico microbiológico de las infecciones de los dispositivos de estimulación cardiaca. Abstract: The use of cardiac implantable electronic devices is rapidly increasing in last years, particularly in older patients and those with significant comorbidities, leading to a higher risk of complications, including infection. In this regard, the detection and identification of the causative microorganisms is crucial for an optimal antimicrobial treatment. In this review, we present the most recent clinical findings about the etiology and microbiological diagnosis of cardiac implantable electronic devices infections.

BackgroundThe translation of Next-Generation Sequencing (NGS) from research to clinical microbiology is increasing rapidly, but its integration into routine clinical care struggles to catch-up. A challenge for clinical laboratories is that the substantial investments made in the required technologies and resources must meet both current and forthcoming needs.MethodsTo get a clinical perspective of these needs, we have sent a survey to infectious diseases clinicians of five hospitals, covering the following topics: NGS knowledge, expected syndromes and patients foreseen to benefit from NGS, and expected impact on antimicrobial prescription.ResultsAccording to clinicians, benefits of NGS are mostly expected in neurological and respiratory infections diagnostics.ConclusionA better dialog between microbiologists and clinicians about hopes and limits of NGS in microbiology may help identifying key investments needed for clinical laboratories, today and tomorrow.

Copyright of Revista de Ciencias Médicas de Pinar del Río is the property of Editorial Ciencias Medicas and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Belonging to the normal oral, gastrointestinal, and urogenital flora, Abiotrophia defectiva is responsible for 1–2 % of all infective endocarditis (IE) cases. The manifestation of A.defectiva endocarditis may by atypical, without fever. Difficult to isolate pathogen requires special culture media. A 45-year-old female was admitted due to anemia and progressive weight loss (8 kg in 6 months). She had a history of benign mitral valve (MV) prolapse and non-stenotic bicuspid aortic valve (BAV). In echocardiography, large vegetations on MV and small vegetation on BAV were found. An enriched medium for fastidious pathogens was used. A. defectiva was identified using biochemical analysis with VITEK-2 Compact. In the fourth week of antibiotic therapy, she required urgent MV replacement due to MV regurgitation progression while vegetation on BAV disappeared. Although patient's frailty and underweight caused prolonged postoperative wound healing, she was transferred to rehabilitation in good conditions. No relapse of IE was observed during five-month follow-up.

Les ostéites du pied chez les patients vivant avec un diabète sucré sont un réel problème de santé publique du fait de leur fréquence dès l'apparition de plaies du pied et de leur évolution dramatique (amputation, décès). L'infection est secondaire à l'atteinte de l'os par de nombreuses bactéries, commensales et pathogènes, qui s'organisent en biofilm autour de la matrice osseuse. Toutefois, peu de bactéries arrivent à pénétrer dans l'os compact. Celles qui y arrivent sont internalisées par les ostéoblastes et elles vont agir sur les ostéoclastes modifiant l'homéostasie de l'os. Le microbiote osseux est particulièrement varié, marqué par un polymicro- bisme et par la forte présence des bactéries anaérobies et du microbiote commensal cutané. Le diagnostic passe par l'analyse microbiologique d'une biopsie osseuse, prélèvement de référence, à réaliser après un débridement de la plaie. L'interprétation des résultats et la prise en charge nécessitent des réunions de concertation pluridisciplinaire. Même si l'utilisation de la biologie moléculaire n'est pas recommandée dans cette pathologie, ces outils permettront sûrement dans l'avenir de raccourcir les délais diagnostiques pour améliorer la prise en charge des patients. Diabetic foot osteomyelitis is a real public health problem.This pathology is frequent when foot ulcers appear and has dramatic consequences with a high risk of amputation and a poor prognosis for the patient with high mortality. The infection is secondary to damage to the bone by numerous bacteria, commensals and pathogens, which are organized into a pathologic biofilm around the bone matrix. However, few bacteria are able to penetrate compact bone. Those that do are internalized by the osteoblasts and act on the osteoclasts modifying the bone homeostasis. The bone microbiota is particularly diverse marked by polymicrobism, the strong presence of anaerobes and bacteria from commensal skin microbiota. The microbiological diagnosis involves a bone biopsy, the reference method, to be performed after wound debridement. The management of the microbiological results frequently requires a multidisciplinary approach. Even if the use of molecular biology is not recommended in this pathology, these tools will surely allow in the future to shorten diagnostic times and improve patient care. [ABSTRACT FROM AUTHOR]

Objective: To describe the use and quality of point-of-care (POC) microscopy, urine culture and susceptibility testing performed in general practice in Northern Denmark from 2013 to 2018. Design: Descriptive study Setting: General practices receiving a fee for examining urine samples. Subjects: Simulated urine samples containing uropathogenic bacteria distributed by the organisation for improvement of microbiological quality (MIKAP). Main outcome measures: Percentage of use and correct answers for microscopy, culture and susceptibility testing. Results: A total of 5361 samples were analysed by the use of microscopy (39.7%), culture (66.0%) and/or susceptibility testing (76.5%). For culture, Flexicult SSI urinary kittm (87.6%) demonstrated the highest percentage of correct answers followed by chromogenic agar (85.1%) and 2-plate dipslide (85.2%). Mueller Hinton agar with tablets had the highest percentage of correct answers for susceptibility testing of most bacterial strains (84.6%), followed by Flexicult (77.2%). Furthermore, susceptibility testing with tablets (range: 76.1–84.6%) was found to be more accurate than discs (range: 72.9–75.5%). Overall, the highest percentage of correct answers was obtained when examining urine samples containing Escherichia coli: Microscopy (78.3%), culture (87.0%) and susceptibility testing (range: 84.3–90.7%). Conclusion: The quality of POC testing in general practice was high when examining urine samples containing the most common uropathogen E. coli. Surprisingly, susceptibility testing was more frequently used than culture. This approach may compromise the treatment decision as only cultures contribute with information about the flora composition and bacterial quantification. Interestingly, microscopy was the least used method even though the result may be reached within a few minutes.Key points The quality of POC tests (microscopy, urine culture, susceptibility testing) performed in general practice was high when examining urine containing E. coli, whereas difficulties were observed for samples including S. saprophyticus or K. pneumoniae. Susceptibility testing was more often performed than urine culture, which indicates a problem as only urine cultures contribute with information about the flora composition and bacterial quantification.

Introduction Actinotignum schaalii is a small Gram-positive facultative anaerobic coccoid rod that was reclassified in 1997 from the genus Actinomyces and is difficult to culture with usual microbiological techniques, as it is slowly growing. A. schaalii is an emerging human pathogen that is most commonly implicated in urinary tract infections (UTIs), but has also been isolated less frequently from abscesses of various sites (such as the skin, the genitourinary tract, surgical sites or intraabdominal). Methods All cases where A. schaalii was identified during a 6-year period (January 2016 - January 2022) in the University Hospital of Heraklion were reviewed. Results A. schaalii was isolated in 11 cultures from 10 patients. The site of infection was skin and soft tissue in nine out of eleven cultures (81.8%) and the bone was the site of infection in two patients (18.2%). Most cultures were polymicrobial. The median age of patients was 55.5 years (interquartile range 34-63 years), and 80% (n=8) were male. A. schaalii was susceptible to beta-lactams, quinolones and vancomycin, but resistant to clindamycin, erythromycin and metronidazole. Moreover, two cases of diabetic patients diagnosed with polymicrobial diabetic foot osteomyelitis from this organism are presented in detail. Both patients were successfully managed with targeted antimicrobial treatment and prompt surgical management. Conclusions A. schaalii is an emerging pathogen that is likely under-reported due to difficulties in isolation and identification. Herein two cases of diabetic foot osteomyelitis are also presented in detail, successfully managed with targeted antimicrobial therapy and surgical debridement. [ABSTRACT FROM AUTHOR]

Background: In the past, various efforts have been made to investigate diagnostic tools for periprosthetic-joint-infection (PJI). It is little-known about the diagnostic utility of polymerase-chain-reaction (PCR) in this context, especially concerning the role of multiplex-PCR assays comparing with conventional tissue culture.Objective: Evaluation of an automated-multiplex-PCR cartridge system for patients with suspicion of PJI in comparison with conventional microbiological culture and 16S-rDNA-PCR.Methods: On suspicion of PJI synovial fluid specimen were taken preoperatively or periprosthetic tissue was collected intraoperatively. Microbiological analysis included conventional culture, 16S-rDNA-PCR and automated-multiplex-PCR (Unyvero-i60-ITI®). The European-Bone-and-Joint-Infection-Society (EBJIS) criteria were used for PJI diagnosis. Positive and negative percent agreement was calculated. Total percentage agreement and Cohen's kappa coefficient were calculated. Sensitivity, specificity and positive predictive value of conventional culture, 16S-rDNA-PCR and multiplex-PCR were calculated. Ten specimens of proved PJI were used as control group.Results: Fifty specimen were suitable for culture. 14 (28%) were classified as PJI, 36 (72%) were aseptic. Coagulase-negative staphylococci was the most frequent detected pathogen. Concordance-rate between mPCR and culture results was 75.6% with a Cohen's kappa of 0.28. Concordance-rate between mPCR and 16S-rDNA was 82.9%, Cohen's kappa was 0.13. Concordance analysis between culture results and 16S-rDNA lead to a concordance-rate of 88.9%. Cohen's kappa was calculated with 0.6. With regard to the microbiological culture as reference, sensitivity of the mPCR was 0.33 and specificity was 0.91. Sensitivity and specificity of the 16S-rDNA-PCR was 0.55 and 0.97. The positive predictive value was 0.57 for the mPCR and 0.83 for the 16S-rDNA-PCR.Conclusions: Due to fair agreement between mPCR and conventional microbiological culture, the tested multiplex-PCR could be an additional instrument for the detection of PJI but is not superior over the conventional culture. [ABSTRACT FROM AUTHOR]

Rat bite fever is characterized by a clinical triad of symptoms, fever, rash and arthritis. It is transmitted by rodents and mainly due to infection by Streptobacillus moniliformis , a fastidious bacterium carried by Rattus norvegicus . This case report presents the case of a patient who developed septic arthritis and fever after a wild rat bite, with subsequent isolation of S. moniliformis from the joint fluid. Upon reviewing 45 other published case reports of S. moniliformis osteoarticular infections following contact with either a rat or its secretions, it was firstly observed that the rat bite fever clinical triad was incomplete in over half of the cases, mainly because rash was infrequently observed among adult patients. Secondly, the clinical presentation of rat bite fever is quite non-specific and rodent exposure is not mentioned by patients in a third of cases upon admission. Altogether, diagnosing rat bite fever is a significant clinical challenge suggesting that it might be significantly underdiagnosed. In addition to these clinical aspects, no evidence was found supporting immunological mechanisms, as suggested in some literature. Instead, when excluding five improperly performed cultures, S. moniliformis was cultured in 25 reported cases and identified twice by direct PCR sequencing amounting to a detection rate of 90% ( n = 27 / 30 ) on joint fluids. Cultures should be performed in medium containing yeast extract, complete peptic digest of animal tissue and at least 5% blood. Knowing that S. moniliformis is very sensitive to many antibiotics thereby making the culture negative, direct 16S rRNA gene sequencing on joint fluid is an alternative method in the case of clinical and cytological evidence of osteoarticular infections with sterile culture of joint fluid., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Giraudon, Larranaga Lapique, Wallemacq, Dalborgo, Yin, Hites and Martiny.)

Abstract Background Leishmaniasis, considered by the World Health Organization as one of the most important tropical diseases, is endemic in the Mediterranean Basin. The aim of this study was to evaluate epidemiological and clinical characteristics of cutaneous (CL) and mucocutaneous leishmaniasis (MCL) in La Fe University Hospital, Valencia, Spain. The particular focus was on diagnosis techniques and clinical differences according to the immunological status of the patients. Methods An eleven-year retrospective observational study of CL and MCL episodes at the hospital was performed. Epidemiological, clinical and therapeutic variables of each case, together with the microbiological and anatomopathological diagnosis, were analyzed. Results A total of 42 patients were included, 30 of them were male and 28 were immunocompetent. Most of the cases (36/42) were diagnosed in the last 5 years (2013–2017). The incidence of CL and MCL increased from 3.6/100,000 (2006–2012) to 13.58/100,000 (2013–2017). The majority of the patients (37/42) exhibited CL, in 30 cases as single lesions (30/37). Ulcerative lesions were more common in immunosuppressed patients (13/14) than in immunocompetent patients (20/28), (P = 0.2302). The length of lesion presence before diagnosis was 7.36 ± 6.72 months in immunocompetent patients and 8.79 ± 6.9 months in immunosuppressed patients (P = 0.1863). Leishmania DNA detection (92.3%) was the most sensitive diagnostic technique followed by Giemsa stain (65%) and histopathological examination (53.8%). Twelve patients (12/42) had close contact with dogs or were living near to kennels, and 10 of them did not present underlying conditions. Intralesional glucantime (21/42) and liposomal amphotericin B (7/42) were the most common treatments administered in monotherapy. All patients evolved successfully and no relapse was reported. Conclusions Some interesting clinical and epidemiological differences were found in our series between immunocompetent and immunosuppressed patients. Future studies can take these results further especially by studying patients with biological therapy. Skin biopsies combining NAAT with histological techniques are the most productive techniques for CL or MCL diagnosis.

Trichomonas vaginalis and Mycoplasma hominis, two microorganisms causing infections of the urogenital tract, are closely associated in that they establish an endosymbiosis relationship, the only case among human pathogens. As a result, the presence of one microorganism may be considered a sign that the other is present as well. Identification of the two pathogens in clinical samples is based on cultivation techniques on specific media, even though in recent years, new sensitive and rapid molecular techniques have become. Here, we demonstrate that the concomitant presence of T.vaginalis in urogenital swabs may lead to a delay in the identification of M.hominis, and thus to an underestimation of bacterial infections when cultural techniques are used.

The global pandemic of COVID-19 has had negative repercussions on the activities and research in clinical microbiology laboratories other than those related to SARS-CoV-2. Nonetheless, the research activity has also continued in other fields. In this brief review, some of the recent publications related to new diagnostic tests, methods for rapid antimicrobial susceptibility testing and for the detection of resistance genes, new diagnostic technologies, and some aspects related to old and emergent pathogens (Candida auris, Elizabethkingia spp. Streptococcus pyogenes) are summarized. [ABSTRACT FROM AUTHOR]

Copyright of Polish Otorhinolaryngological Review / Polski Przegląd Otorynolaryngologiczny (Index Copernicus) is the property of Index Copernicus International and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Corynebacterium diphtheriae is a gram-positive bacillus with characteristic polar metachromatic granules. It was a significant public health problem, infecting the throat and upper airways and producing a highly potent exotoxin but with the advent of diphtheria vaccine, cases have rapidly decreased. Now over the last decade, the reemergence of this infection has been noted and case reports from India have been documented. India represents 78% of globally reported cases with significant mortality despite national immunization programs in place. This case study indicates the severity of an improperly managed case, the importance of microbiological diagnosis with a special interest in molecular detection, and reinforces a resurgence of diphtheria infection.

Abstract Background Data on the methods used for microbiological diagnosis of hospital-acquired pneumonia (HAP) are mainly extrapolated from ventilator-associated pneumonia. HAP poses additional challenges for respiratory sampling, and the utility of sputum or distal sampling in HAP has not been comprehensively evaluated, particularly in HAP admitted to the ICU. Methods We analyzed 200 patients with HAP from six ICUs in a teaching hospital in Barcelona, Spain. The respiratory sampling methods used were divided into non-invasive [sputum and endotracheal aspirate (EAT)] and invasive [fiberoptic-bronchoscopy aspirate (FBAS), and bronchoalveolar lavage (BAL)]. Results A median of three diagnostic methods were applied [range 2–4]. At least one respiratory sampling method was applied in 93% of patients, and two or more were applied in 40%. Microbiological diagnosis was achieved in 99 (50%) patients, 69 (70%) by only one method (42% FBAS, 23% EAT, 15% sputum, 9% BAL, 7% blood culture, and 4% urinary antigen). Seventy-eight (39%) patients underwent a fiberoptic-bronchoscopy when not receiving mechanical ventilation. Higher rates of microbiological diagnosis were observed in the invasive group (56 vs. 39%, p = 0.018). Patients with microbiological diagnosis more frequently presented changes in their empirical antibiotic scheme, mainly de-escalation. Conclusions A comprehensive approach might be undertaken for microbiological diagnosis in critically ill nonventilated HAP. Sputum sampling determined one third of microbiological diagnosis in HAP patients who were not subsequently intubated. Invasive methods were associated with higher rates of microbiological diagnosis.

Background: In prosthetic joint infections (PJIs), identification of the causative microorganisms is critical to successfully adapt and optimize treatment. However, microbiological diagnosis of PJIs remains a challenge notably because bacteria are embedded in biofilm adhered to the prosthetic material. Recently, dithiothreitol (DTT) treatment of prosthesis has been proposed as a new strategy to release bacteria from biofilm and to improve the yield of microbiological diagnosis. In this study, we evaluated the interest of a commercial device using DTT, the MicroDTTect system (Heraeus, Hanau, Germany), for the diagnosis of low-grade chronic PJIs, compared to the conventional culture of periprosthetic tissue (PPT) samples.Methods: Twenty patients undergoing a surgery procedure for removal of prosthetic material because of a suspicion of low-grade PJI without pre-operative microbiological documentation were included (NCT04371068). Bacteriological results using the fluid obtained after prosthesis treatment with the MicroDTTect system were compared to results obtained with conventional culture of PPT samples.Results: All the bacteria considered as responsible for PJIs recovered from culture of PPT samples were also detected using the MicroDTTect device. For one patient, an additional bacterial isolate (Staphylococcus haemolyticus) suspected to be involved in a polymicrobial PJI was identified using DTT treatment. Time to positivity of the cultures was also reduced using the MicroDTTect system, notably in case of Cutibacterium acnes infection. However, probable bacterial contaminants were found (MicroDTTect system, n = 5; PPT samples, n = 1).Conclusion: This study showed that DTT treatment of the prosthetic component using the MicroDTTect device could improve the microbiological diagnosis of low-grade PJIs.

Background: Intramedullary nailing (IMN) has been frequently indicated to treat long bone open and closed fractures, but IMN infection (IMNI) may have devastating consequences. Sonication has been regarded as an important add-on for microbial identification on a variety of orthopaedic implant-associated infections, but its role in the IMNI is poorly studied. We aim at evaluating the accuracy obtained by conventional peri-implant tissue culture (TC) samples with sonication fluid cultures (SCs) of IMNI.Methods: Longitudinal prospective cohort study ongoing since June 2014, which included patients with indication for IMN removal due to any reason. Clinical diagnosis of INMI was defined according to publication addressing fracture-related infections. Minimal of two samples from TC were cultured. SCs followed the protocol previously published. Statistical analysis was performed using McNemar's test for related proportions.Results: We included 54 patients submitted to IMN retrieval, of whom 47 presenting clinical signs of IMNI. Sensitivity for detecting microorganisms using TC and SC was 89.4% (42/47) and 97.6% (40/41), and specificity was 71.4% (5/7) for both TC and SC (p = 1.00). Positive and negative predictive values for TC and SC were 95.5% (42/44), 95.2% (40/42), 50% (5/10), and 83.3% (5/6), respectively. The most frequent organisms isolated in both TC and SC were Staphylococcus aureus, S. epidermidis, and Enterococcus sp. Polymicrobial infection was diagnosed in 14.8% (8/54) and 25% (12/48) by TC and SC, respectively (p = 0.19).Conclusion: Sonication fluid and tissue samples presented similar accuracy on the diagnosis of IMNI, but SC was advantageous of detecting polymicrobial infection. [ABSTRACT FROM AUTHOR]

Because the occurrence of infective endocarditis (IE) continues to be associated with high mortality, a working group was created by the Dutch Society of Cardiology to examine how the most recent European Society of Cardiology (ESC) guidelines for IE management could be implemented most effectively in the Netherlands. In order to investigate current Dutch IE practices, the working group conducted a country-wide survey. Based on the results obtained, it was concluded that most ESC recommendations could be endorsed, albeit with some adjustments. For instance, the suggested pre-operative screening and treatment of nasal carriers of Staphylococcus aureus as formulated in the ESC guideline was found to be dissimilar to current Dutch practice, and was therefore made less restrictive. The recently adapted ESC diagnostic criteria for IE were endorsed, while the practical employment of the relevant diagnostic techniques was simplified in an adapted flowchart. In addition, the presence of a multidisciplinary, so-called 'endocarditis team' in tertiary centres was proposed as a quality indicator. An adapted flowchart specifically tailored to Dutch practice for microbiological diagnostic purposes was constructed. Lastly, the working group recommended the Stichting Werkgroep Antibioticabeleid (SWAB; Dutch Working Party on Antibiotic Policy) guidelines for IE treatment instead of the antibiotic regimens proposed by the ESC. [ABSTRACT FROM AUTHOR]

Vertebral osteomyelitis (VOM) is often diagnosed with delays, resulting in poorer outcomes. Microbial documentation is particularly challenging and obtained using blood cultures (BCs) and vertebral biopsies (VBs; CT-guided or surgical). We retrospectively analysed VOM cases in a tertiary reference centre between 2004 and 2015, focusing on how and how quickly microbiological diagnosis was performed. Among 220 VOM, 88.2% had documentation, including Gram-positive cocci (GPC) (70.6%), Gram-negative rods (GNR) (9.3%), anaerobes (3.6%), polybacterial infections (6.7%) and tuberculosis (9.8%). BCs were performed in 98.2% and positive in 59.3%, identifying most GPC (80.3%) and half of GNR (54.6%). VBs were performed in fewer cases (37.7%), but were more frequently positive (68.8% for CT-guided and 81.0% for surgical biopsies). They documented all anaerobes (100.0%), most M. tuberculosis (84.2%) and polybacterial infections (76.9%), and GNR (45.4%). Extra-vertebral samples highly contributed to tuberculosis diagnosis (52.6%, and 15.8% as the only positive sample). Documentations most often followed radiological diagnosis (53.4%). They were obtained earlier by BCs than by VB after first clinical symptoms (median of 14 versus 51 days). Antibiotic treatments were mostly initiated after samplings (88.0%). BCs allow the documentation of most VOM and should be performed without delay in case of clinical or radiological suspicion; however, they may miss 1 out of 5 GPC and 1 out of 2 GNR. VBs have a higher positivity rate and should be rapidly performed if negative BCs. It is likely that delayed and missed diagnoses result from the insufficient use of VB. [ABSTRACT FROM AUTHOR]

Background: The potential differences between a clinical diagnosis of coronavirus disease 2019 (COVID-19) (i.e., symptoms without positive virus test) and a microbiological diagnosis (i.e., positive virus test results) of COVID-19 are not known. Aims: This study explored the differences between the two types of COVID-19 diagnosis among older patients in terms of clinical characteristics and outcomes. Methods: A total of 244 inpatients aged ≥ 60 years with COVID-19 were included in this study, of whom 52 were clinically diagnosed and 192 were microbiologically diagnosed. Clinical and laboratory data on hospital admission and outcomes (discharged or died in hospital) of all patients were retrieved from medical records retrospectively. Patients who met the criteria for clinical diagnosis with negative virus test results were assigned to the clinical diagnosis group, whereas those with positive virus test results were assigned to the microbiological diagnosis group. After univariate analyses, two propensity score analyses [i.e., covariate adjustment using propensity score (CAPS) and propensity score matching (PSM)] were conducted to control bias. Results: The clinical and microbiological diagnosis groups demonstrated significant differences in outcomes and in the majority of laboratory findings. After propensity score analyses, many differences between the two groups disappeared and the rate of mortality had no statistically significant difference (P = 0.318 and 0.828 for CAPS and PSM, respectively). Conclusions: Patients with similar signs, symptoms, and laboratory and imaging findings as confirmed COVID-19 cases may have a similar mortality risk, regardless of the virus test results, and require timely intervention to reduce their mortality. [ABSTRACT FROM AUTHOR]

World Health Organization, named the disease caused by Severe Acute Respiratory syndrome-coronavirus-2 coronavirus on 11 February 2020 as Coronavirus Disease-2019 (COVID-19). A person who has clinical symptom and a history of contact with a patient with confirmed COVID-19 is identified as a ''possible case'' and is expected to be confirmed by laboratory test. A confirmed case is asymptomatic/symptomatic suspected individual with a positive molecular test. Specific diagnosis is made by specific molecular tests on respiratory samples (orafarengeal/ nasopharyngeal swab/sputum/endotracheal aspirates and bronchoalveolar lavage). The virus can also be detected in the stool and in severe patients for a short time in the blood. The number of molecular tests that should be used in the case of a pandemic is rapidly increasing. At the beginning of the outbreak in Turkey, tests could only be performed in suspected patients in the central unit and then they began to be performed in other reference laboratories. Rapid antigen tests did not provide the desired performance, while rapid antibody tests were distributed to the field to support polymerase chain reaction tests and determine immunity. Microbiology Laboratory is indispensable in controlling the COVID-19 pandemic. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis, considered by the World Health Organization as one of the most important tropical diseases, is endemic in the Mediterranean Basin. The aim of this study was to evaluate epidemiological and clinical characteristics of cutaneous (CL) and mucocutaneous leishmaniasis (MCL) in La Fe University Hospital, Valencia, Spain. The particular focus was on diagnosis techniques and clinical differences according to the immunological status of the patients. Methods: An eleven-year retrospective observational study of CL and MCL episodes at the hospital was performed. Epidemiological, clinical and therapeutic variables of each case, together with the microbiological and anatomopathological diagnosis, were analyzed. Results: A total of 42 patients were included, 30 of them were male and 28 were immunocompetent. Most of the cases (36/42) were diagnosed in the last 5 years (2013–2017). The incidence of CL and MCL increased from 3.6/100,000 (2006–2012) to 13.58/100,000 (2013–2017). The majority of the patients (37/42) exhibited CL, in 30 cases as single lesions (30/37). Ulcerative lesions were more common in immunosuppressed patients (13/14) than in immunocompetent patients (20/28), (P = 0.2302). The length of lesion presence before diagnosis was 7.36 ± 6.72 months in immunocompetent patients and 8.79 ± 6.9 months in immunosuppressed patients (P = 0.1863). Leishmania DNA detection (92.3%) was the most sensitive diagnostic technique followed by Giemsa stain (65%) and histopathological examination (53.8%). Twelve patients (12/42) had close contact with dogs or were living near to kennels, and 10 of them did not present underlying conditions. Intralesional glucantime (21/42) and liposomal amphotericin B (7/42) were the most common treatments administered in monotherapy. All patients evolved successfully and no relapse was reported. Conclusions: Some interesting clinical and epidemiological differences were found in our series between immunocompetent and immunosuppressed patients. Future studies can take these results further especially by studying patients with biological therapy. Skin biopsies combining NAAT with histological techniques are the most productive techniques for CL or MCL diagnosis. [ABSTRACT FROM AUTHOR]

Copyright of Acta Facultatis Medicae Naissensis is the property of Nis University, Faculty of Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To assess the clinical accuracy (sensitivity (SEN), specificity (SPE), positive predictive value and negative predictive value) of two point-of-care (POC) urine culture tests for the identification of urinary tract infection (UTI) in general practice. Design: Prospective diagnostic accuracy study comparing two index tests (Flexicult™ SSI-Urinary Kit or ID Flexicult™) with a reference standard (urine culture performed in the microbiological department). Setting: General practice in the Copenhagen area patients. Adult female patients consulting their general practitioner with suspected uncomplicated, symptomatic UTI. Main outcome measures: (1) Overall accuracy of POC urine culture in general practice. (2) Individual accuracy of each of the two POC tests in this study. (3) Accuracy of POC urine culture in general practice with enterococci excluded, since enterococci are known to multiply in boric acid used for transportation for the reference standard. (4) Accuracy based on expert reading of photographs of POC urine cultures performed in general practice. Standard culture performed in the microbiological department was used as reference standard for all four measures. Results: Twenty general practices recruited 341 patients with suspected uncomplicated UTI. The overall agreement between index test and reference was 0.76 (CI: 0.71–0.80), SEN 0.88 (CI: 0.83–0.92) and SPE 0.55 (CI: 0.46–0.64). The two POC tests produced similar results individually. Overall agreement with enterococci excluded was 0.82 (0.77–0.86) and agreement between expert readings of photographs and reference results was 0.81 (CI: 0.76–0.85). Conclusions: POC culture used in general practice has high SEN but low SPE. Low SPE could be due to both misinterpretation in general practice and an imperfect reference standard. Registration number: ClinicalTrials.gov NCT02323087.

Nosocomial pneumonia is a healthcare-associated infection with significant consequences for the patient and the healthcare system. The efficacy of treatment significantly depends on the timeliness and adequacy of the antibiotic therapy regimen. The growth of resistance of gram-negative pathogens of nosocomial pneumonia to antimicrobial agents increases the risk of prescribing inadequate empirical therapy, which worsens the results of patient treatment. Identification of risk factors for infection with multidrug-resistant microorganisms, careful local microbiological monitoring with detection of resistance mechanisms, implementation of antimicrobial therapy control strategy and use of rational combinations of antibacterial drugs are of great importance. In addition, the importance of using new drugs with activity against carbapenem-resistant strains, including ceftazidime/aviabactam, must be understood. This review outlines the current data on the etiology, features of diagnosis and antibacterial therapy of nosocomial pneumonia.

Abstract Background Encephalitis and meningoencephalitis are severe, sometime life-threatening infections of the central nervous system. Travellers may be exposed to a variety of neurotropic pathogens. Aims We propose to review known infectious causes of encephalitis in adults acquired outside Europe, and how to identify them. Sources We used Pubmed and Embase, to search the most relevant publications over the last years. Content Microbiologic tests and radiological tools to best identify the causative pathogen in travellers presenting with encephalitis and ME are presented in this narrative review, as well as a diagnostic approach tailored to the visited area and types of exposures. Implications This review highlights the diagnostic difficulties inherent to exotic causes of central nervous system infections, and attempts to guide clinicians with respect to which microbiological tests to consider, in addition to brain MRI, when approaching a returning traveller presenting with encephalitis. [ABSTRACT FROM AUTHOR]

Community-acquired pneumonia (CAP) is one of the most common causes of hospital admissions and death worldwide. The incidence and mortality of CAP are associated with the presence of comorbidities and increasing age. Streptococcus pneumoniae is the most frequent causative microorganism of CAP, although in many patients with CAP, the causative microorganism remains unknown. Currently, antimicrobial resistance is increasing, so the accurate diagnosis and determination of the causative microorganism are even more important. This is a key point in reducing both morbidity and mortality from CAP, and appropriate antimicrobial stewardship is now a global priority. This review summarizes on the epidemiology, microbiological etiology, and diagnosis of CAP in adults. [ABSTRACT FROM AUTHOR]

Post-mortem diagnosis of sepsis is often very difficult to make, especially in the elderly affected by multiple comorbidities. However, clinical evaluation following histology, immunohistochemistry, microbiological tests, immunoassays and proteomics can improve reliability of this post-mortem diagnosis. [ABSTRACT FROM AUTHOR]

Corynebacterium diphtheriae is a gram-positive bacillus with characteristic polar metachromatic granules. It was a significant public health problem, infecting the throat and upper airways and producing a highly potent exotoxin but with the advent of diphtheria vaccine, cases have rapidly decreased. Now over the last decade, the reemergence of this infection has been noted and case reports from India have been documented. India represents 78% of globally reported cases with significant mortality despite national immunization programs in place. This case study indicates the severity of an improperly managed case, the importance of microbiological diagnosis with a special interest in molecular detection, and reinforces a resurgence of diphtheria infection. [ABSTRACT FROM AUTHOR]

Aggregatibacter actinomycetemcomitans is a key pathogen that has been associated with periodontal disease. Its most important virulence factor is a leukotoxin capable of inactivating immune cells. The JP2 genotype of Aggregatibacter actinomycetemcomitans shows enhanced leukotoxic activity and is mostly present in individuals of North and West African origin with severe periodontitis. In this paper, two cases of Caucasians diagnosed with the JP2 genotype are presented. A 50-year-old female patient had three approximal sites with ≥ 6 mm clinical attachment loss (CAL) and eight sites with probing depth (PD) ≥ 5 mm. Microbiological diagnostics revealed A. actinomycetemcomitans JP2 genotype, but not Porphyromonas gingivalis. This JP2 genotype was highly leukotoxic to monocytic cells. The second case was a 55-year-old female patient with CAL of > 5 mm at all molars and PD of up to 12 mm. A. actinomycetemcomitans JP2 was identified, but not P. gingivalis. Her husband originated from North-Africa. In him, no A. actinomycetemcomitans was detected, but their 17-year-old daughter was diagnosed with periodontitis and was found to be positive for the JP2 genotype. Both patients were successfully treated with adjunctive antibiotics and the JP2 genotype was eliminated. In summary, here, the microbiological diagnosis was key for the treatment with adjunctive antibiotics.

Background: Vascular graft infections (VGI) are difficult to diagnose and treat, and despite redo surgery combined with antimicrobial treatment, outcomes are often poor. VGI diagnosis is based on a combination of clinical, radiological, laboratory and microbiological criteria. However, as many of the VGI patients are already under antimicrobial treatment at the time of redo surgery, microbiological identification is often difficult and bacterial cultures often remain negative rendering targeted treatment impossible. We aimed to assess the benefit of 16S rRNA gene polymerase chain reaction (broad-range PCR) for better microbiological identification in patients with VGI.Methods: We prospectively analyzed the clinical, microbiological, and treatment data of patients enrolled in the observational Vascular Graft Cohort Study (VASGRA), University Hospital Zurich, Switzerland. The routine diagnostic work-up involved microbiological cultures of minced tissue samples, and the use of molecular techniques in parallel. Patient-related and microbiological data were assessed in descriptive analyses, and we calculated sensitivity, specificity, negative and positive predictive value for broad-range 16S rRNA gene PCR versus culture (considered as gold standard).Results: We investigated 60 patients (median age 66 years (Interquartile range [IQR] 59–75)) with confirmed VGI between May 2013 and July 2017. The prevalence of antimicrobial pretreatment at the time of sampling was high [91%; median days of antibiotics 7 days (IQR 1–18)]. We investigated 226 microbiological specimens. Thereof, 176 (78%) were culture-negative and 50 (22%) were culture-positive. There was a concordance of 70% (158/226) between conventional culture and broad-range PCR (sensitivity 58% (95% CI 43–72); specificity 74% (67–80%)). Among the group of 176 culture-negative specimens, 46 specimens were broad-range PCR-positive resulting in identification of overall 69 species. Among the culture and/or broad-range PCR-positive specimens (n = 96), 74 (77%) were monomicrobial and 22 (23%) polymicrobial, whereas the rate of polymicrobial samples was higher in broad-range PCR-positive specimens (93%).Conclusions: Combined cultures and broad-range 16S rRNA gene PCR from periprosthetic tissue and/or explanted vascular grafts increased the diagnostic accuracy in VGI, particularly in patients already under antimicrobial treatment at the time of redo surgery. Ideally, antimicrobial treatment should be withheld until surgical sampling in order to optimize microbiological diagnostics.Clinical trials.gov identifier: NCT01821664