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Anti- Aspergillus IgG antibodies are important biomarkers for the diagnosis of chronic pulmonary aspergillosis (CPA) and allergic bronchopulmonary aspergillosis (ABPA). We compared the performance of a new commercial enzyme immunoassay (EIA) (Bordier Affinity Products) with that of the Bio-Rad and Virion\Serion EIAs. This assay is novel in its association of two recombinant antigens with somatic and metabolic antigens of Aspergillus fumigatus . In a prospective multicenter study, 436 serum samples from 147 patients diagnosed with CPA (136 samples/104 patients) or ABPA (94 samples/43 patients) and from 205 controls (206 samples) were tested. We obtained sensitivities of 97%, 91.7%, and 86.1%, and specificities of 90.3%, 91.3%, and 81.5% for the Bordier, Bio-Rad, and Virion\Serion tests, respectively. The Bordier kit was more sensitive than the Bio-Rad kit ( P < 0.01), which was itself more sensitive than the Virion\Serion kit ( P = 0.04). The Bordier and Bio-Rad kits had similar specificity ( P = 0.8), both higher than that of the Virion\Serion kit ( P = 0.02). The area under the receiver operating characteristic (ROC) curves confirmed the superiority of the Bordier kit over the Bio-Rad and the Virion\Serion kits (0.977, 0.951, and 0.897, respectively; P < 0.01 for each comparison). In a subset analysis of 279 serum samples tested with the Bordier and Bio-Rad kits and an in-house immunoprecipitin assay (IPD), the Bordier kit had the highest sensitivity (97.7%), but the IPD tended to be more specific (71.2 and 84.7%, respectively; P = 0.10). The use of recombinant, somatic, and metabolic antigens in a single EIA improved the balance of sensitivity and specificity, resulting in an assay highly suitable for use in the diagnosis of chronic and allergic aspergillosis.

International audience; BackgroundFew studies have suggested the potential role of respiratory viruses in cystic fibrosis (CF) exacerbation, but their real impact is probably underestimated.MethodSixty-four sputum samples collected from 46 adult patients were included in the study: 33 samples were collected during exacerbation of CF, and 31 during the stable phase. After extraction, nucleic acids were tested for the presence of respiratory viruses. When rhinovirus (HRV) was detected, the 5′UTR and VP4/2 regions were sequenced, and phylogenetically analyzed. The characteristics of patients in exacerbation and stable phase were compared.ResultsViruses were found in 25% of samples. The HRV viruses were the most frequently detected followed by coronaviruses. Only the HRV detection was significantly associated with the occurrence of CF pulmonary exacerbation (p < 0.027). Characterization of 5′UTR and VP4/2 regions of the HRV genome specified that HRV-A, -B, -C were detected. All HRV-C were recombinant HRV-Ca.ConclusionsHRV were the most frequently detected viruses; their detection was significantly associated with the occurrence of an exacerbation. The reality of viral recombination between HRV was demonstrated in CF patients for the first time, raising the role of viruses in lung microbiota. Further studies are now warranted to decipher virus impact in CF

Introduction Our objective was to investigate characteristics risk factors and outcomes of patients with chronic pulmonary aspergillosis (CPA). Methods The Aspergillosis Committee prospectively collected Aspergillus notifications from January 2000 to December 2011. A retrospective analysis of data was performed. Results Among 1614 notifications registered, 44 cases of CPA in non-immunocompromised patients were identified. The median age was 65 years (Q1–Q3: 54–75), the median body mass index (BMI) was 20 kg/m2 (Q1–Q3: 16–22) and 15 had chronic obstructive pulmonary disease. All patients had a positive specific serum precipitin antibody titer. Radiological presentations were: cavitations [single n = 31 (70%); multiple n = 12 (27%)] containing mycetomas [n = 18 (41%)], consolidations [n = 19 (43%)], emphysema [n = 15 (34%)] and sequelae of mycobacterial infection [n = 10 (23%)]. The median duration of follow-up was 30 months (Q1–Q3: 14–55). The median duration of antifungal treatment was 6 months (Q1–Q3: 3–12). Outcomes were unfavorable in 14 patients, and 12 (27%) died. Analysis by multivariate Cox regression model with bootstrapping showed that a higher BMI and a lower Charlson index score were predictive of favorable evolution, hazard ratio (95% confidence interval): BMI (+1) = 0.83 (0.71–0.97), Charlson (+1) = 1.37 (1.01–1.85). When analyses were restricted to chronic CPA and chronic necrotizing pulmonary aspergillosis, the multivariate Cox regression model showed that both BMI and Charlson index score were not statistically significant. Conclusion Our results provide data on clinical characteristics and outcomes of CPA emphasizing the role of preexisting chronic respiratory conditions and protective effect of preserved BMI and lower Charlson index score.

International audience; Borrelia crocidurae-associated relapsing fever is endemic to West Africa and is considered benign. We report 4 patients with B. crocidurae-associated neurologic symptoms; 2 of their cases had been misdiagnosed. Frequency and severity of this disease could be underestimated; molecular methods and serodiagnostic tests for Lyme disease might be helpful in its detection.

Allergic bronchopulmonary aspergillosis (ABPA), which results from hypersensitivity, primarily to Aspergillus, represents a severe complication in patients suffering from asthma or cystic fibrosis (CF). Since early treatment of ABPA is supposed to prevent long-term damages, ABPA has to be diagnosed promptly. However, this diagnosis is not straightforward due to clinical and radiological features of ABPA overlapping with those of CF. Despite ABPA specific diagnosis criteria proposed by the Cystic Fibrosis Foundation in 2003, making a definitive ABPA diagnosis in CF patients remains a challenge. Recent advances in the immunopathogenesis of ABPA have initiated the development of new serological tests, such as the recently reported detection of specific IgE to recombinant A. fumigatus allergens, or Thymus- and activation-regulated chemokine (TARC / CCL17), both of which are of value in the diagnosis of APBA. We review in this paper the serum markers that can advance ABPA diagnosis in CF patients, ranging from the well known criteria (anti-A. fumigatus IgE, IgG, and precipitins) to the recent biomarkers (IgE towards recombinant A. fumigatus allergens or TARC detection). Taking into account the up-dated physiopathology of ABPA, we discuss their place and their usefulness, especially TARC, to improve early ABPA detection and monitoring in CF patients.

We conducted a retrospective study to evaluate the usefulness of immunoglobulin G (IgG) subclasses against Candida cell wall fragments (CW) and phosphopeptidomannan (PPM) for the diagnosis of invasive candidiasis (IC). We analyzed 54 patients with IC (n = 19), Candida heavy colonization (HC; n = 16), and controls (no IC or HC, n = 19).In nonneutropenic patients (n = 47), the sensitivity and specificity values of IgG1 anti-CW and IgG2 anti-PPM in IC were 88%, 59%, and 88%, 94%, respectively. The areas under the receiver operating characteristic curves were 0.69 (0.51-0.88) and 0.901 (0.78-1.02), respectively. IgG1 mean values (arbitrary units) and 95% confidence interval were 46 (20-71), 42 (-0.38 to 84) and 20 (8.3-32) in IC, HC, and in controls, respectively, and discriminated IC but not HC from controls (P = .032, and P = .77, respectively). IgG2 mean values were 26 (9.2-42), 19 (4.4-33), and 3.2 (0.28-6.6) in IC, HC, and in controls, respectively, and discriminated both IC and HC from controls (P < .0001 and P = .035, respectively) but did not separate IC from HC (P = .2). IgG2 showed positivity as early as one day after the IC diagnosis. Antibodies were detected in only two out of a total of seven neutropenic patients.For both IC and HC patients, the diagnostic performance of IgG2 anti-PPM was better than the one of IgG1 anti-CW. In nonneutropenic patients, IgG2 anti-PPM accurately identified not only IC patients but also HC patients at high risk for IC. This marker may help clinicians in the initiation of early preemptive therapy.

In a multicenter study, potassium dichromate-preserved stools from patients infected with Cryptosporidium parvum ( n = 20), C. hominis ( n = 20), and other Cryptosporidium species ( n = 10) and 60 controls were examined using four immunochromatographic assays. Assay sensitivity ranged between 50.1% and 86.7% for C. parvum and C. hominis but was

The gold standard laboratory tests used to diagnose invasive Candida infection (ICI) are based on the in vitro culture of blood or samples from other sterile sites. However, these tests have limited sensitivity (Se) and are generally not diagnostic until late in the infectious process. The Serion Candida mannan kit was evaluated for the diagnosis of ICI at Grenoble University Hospital (France) between 2007 and 2011. The results were then compared with worldwide data published between 1997 and 2011. This retrospective study was based on follow-up from the investigation of 162 patients of whom 91 had proven ICI; 13 had Candida colonization index (CCI) scores ≥0.42, positive mannan tests, with nonconcomitant infections; and 58 had no evidence of Candida infection. Candida albicans, C. glabrata, C. tropicalis, and C. parapsilosis were the etiologic agents in 104 patients. For patients with or without ICI, the 12-week mortality rates were 35/104 (33.7%) and 6/58 (10.3%), respectively. The mannan diagnostic specificity was 51% and Se was 77%. However, in the meta-analysis (n = 1,536), values were 86% and 62%, respectively. Positive mannan test results may appear early (median 6 days) in the development of candidemia and have moderate diagnostic value for ICI, with a negative predictive value of 83%. In patients at risk of ICI with negative candidemia, the combination of Candida mannan test data with a CCI score ≥0.42 may improve the diagnosis of probable ICI.

We have developed an inhibition enzyme immunoassay (inhibition-EIA) to monitor for the occurrence of invasive aspergillosis (IA) in sera from 45 immunocompromised (IC) patients. The test uses rabbit polyclonal antibodies and a mixture of components from Aspergillus fumigatus , containing three predominant antigens with molecular weights of 18,000, 33,000, and 56,000. Circulating antigens were found in five of seven proven cases of IA due to A. fumigatus . In two of the five positive cases, antigenemia was detected with inhibition-EIA earlier than with X ray or other biological methods. No antigens were detected in the sera from two patients with proven IA due to Aspergillus flavus and Aspergillus terreus nor in the sera from four patients with probable IA. Circulating antigens were not detected in the control group, composed of 30 healthy adult blood donors. Four of the 32 at-risk patients examined, though they displayed no definite evidence of IA, gave a positive result in this test. The sensitivity, specificity, and positive predictive value of inhibition-EIA were 71.4, 94.4, and 71.2%, respectively. The data were compared with those obtained by a latex agglutination assay of galactomannan (GM) that was positive in only one patient with probable IA. The higher sensitivity obtained by inhibition-EIA may well be due to its ability to detect circulating antigens other than GM in the sera of IC patients with IA. Detecting these antigens may improve the diagnosis of IA, as they may serve as markers of this infection.

To analyse if hydrocortisone could modify the in vitro susceptibility of Aspergillus fumigatus to antifungal drugs, we developed a procedure to test the susceptibility of an A. fumigatus strain to amphotericin B and itraconazole, grown in the presence and in the absence of hydrocortisone. Conidia were germinated in the presence or the absence of hydrocortisone in Czapek medium without antifungal drug. A dilution of thesecultures (5 × 103 conidia ml-1) was spread onto Czapek-agarose plates containing both antifungal drug and hydrocortisone. The cfu per plate were enumerated and compared. A therapeutic concentration of hydrocortisone induced a significant increase in the susceptibility to itraconazole. Conversely, the susceptibility to amphotericin B was not significantly modified when this antifungal drug was associated with hydrocortisone.

The effectiveness of galactomannan detection with the Platelia test was evaluated in a prospective study of 3,327 sera from 807 patients. The specificity was 99.6% (748 of 751 cases). For the groups of patients with proven and probable invasive aspergillosis, the sensitivity was 50.0% (17 of 34 cases). The disappointing sensitivity associated with the presence of rare false-positive cases underlines the limits of this test.

International audience; OBJECTIVES: This study aims at describing the evolution of the epidemiology of invasive aspergillosis (IA) in a French University Hospital focussing on nosocomial cases, in order to assess the efficiency of the environmental preventive measures which were implemented. METHODS: From 2003 to 2009, IA cases were reviewed monthly and classified according to the EORTC/MSG criteria and the origin of contamination. RESULTS: Five proven and 65 probable IA cases were diagnosed. Most of the cases (74.3%) occurred in patients with haematological malignancies. Incidences of IA and nosocomial IA (NIA) were 0.106 and 0.032 cases per 1000 admissions, respectively. All the 21 NIA cases occurred in the absence of air treatment (laminar air flow facilities or Plasmair decontamination units) and/or during construction works. The 3-month and 1-year overall survival rates were 50.6% [38.2-61.7] and 31.1% [20-42.9] respectively, and did not differ according to the origin of contamination. CONCLUSION: Nosocomial IA still accounted for a third of all IA cases diagnosed from 2003 to 2009 and mainly occurred in the absence of environmental protective measures, which were confirmed to be effective when applied. Our results show that extension and/or reinforcement of these measures is needed, especially in the haematology unit and during construction works.

Eight recombinant proteins and purified galactomannan of Aspergillus fumigatus were tested by enzyme-linked immunosorbent assay to quantify the anti-Aspergillus antibodies in sera of patients with aspergilloma, allergic bronchopulmonary aspergillosis (ABPA), and invasive aspergillosis (IA). In spite of the variability observed in the immune responses of individual patients, quantification of the antibody titers against the 18-kDa ribonuclease (RNU), the 360-kDa catalase (CAT), and the 88-kDa dipeptidylpeptidase V (DPPV) was useful for the diagnosis of aspergilloma and ABPA. Differential diagnosis of ABPA was even possible among cystic fibrosis as well as noncystic fibrosis patients. In the group of immunocompromised patients with IA, no antibody response was mounted in response to the Aspergillus infection in any of the patients. Interestingly, about half of the patients with proven IA came to the hospital with high titers of anti-Aspergillus antibodies, suggesting that they were infected upon entry to the hospital. These results suggest that recombinant RNU, CAT, and DPPV have a great potential in the serodiagnosis of all forms of aspergillosis in the immunocompromised and immunocompetent patient.

QBC, examination of thin blood smears, and Parasight-F were performed for every case of malaria suspected between May 1997 and December 1998. Data from 310 patients were reported. Fifty had malaria infection diagnosed by QBC and thin blood film, among whom 39 had Plasmodium falciparum infection. Three of these 39 were negative with the Parasight-F test. Eleven patients had a positive ParaSight-F test but microscopic diagnosis methods were negative. Interpretation of these 11 positive ParaSight-F results is proposed to depend on clinical criteria.

OBJECTIVE To identify alpha-amylase crystalloid formations in parotid specimens obtained by fine needle aspiration. STUDY DESIGN The study concerned three cases of sialadenitis with crystalloid formation observed between 1993 and 1998. In one of these cases, transmission electron microscopy, mass spectrometry and measurement of amylase activity were used to characterize the nature of the crystalloids. RESULTS Light microscopy revealed the same crystalloid structure in all three cases. In one case, where the material was saved, a biochemical method made it possible to reveal high amylase activity, while protein electrophoresis and mass spectrometry were used to identify salivary alpha-amylase. CONCLUSION Crystalloids of salivary alpha-amylase can be identified by May-Grunwald-Giemsa and Papanicolaou stain and can be rapidly confirmed through determination of amylase activity.

Human cryptosporidiosis : an emerging parasitic disease of increasing importance in public health. Cryptosporidiosis in human is an emerging parasitosis due to an intestinal and opportunistic protozoan : Cryptosporidium parvum. This parasite was described 100 years ago in calves and lambs. This zoonosis had emerged as a global public health problem in industrial countries for essentially two reasons. One is linked to its potentially life threatening evolution which remains practiccaly incurable, in severely immunocompromised patients (AIDS patients, transplant patients, young children with immature immunological system...). The other is the important warterbome outbreaks in industrial countries. Clinical forms of cryptosporidiosis range from asymptomatic to acute intestinal illness in immunocompetent host. In immunocompromised host, chronic or severe diarrhoeal diseases increase the morbidity and mortality. Moreover extra intestinal dissemination of parasites can accured (cholangitis) and pulmonary cryptosporidiosis are also frequently discribed. This parasitosis is also caracterized by the lack of efficient therapy and insuffisant prophylactic measures. Numerical standard and legal regulation of Cryptosporidium was proposed. Furthermore the oocyts are extremelly resistant to conventional disinfectants. The control of water treatment is necessary to determine the efficiency of prophylactic measures. The significance of different Cryptosporidium parvum isolates could suggested the existence of different transmission cycles in human and merits futher investigations. The virulent potential of Cryptosporidium strains as well as individual susceptibility need further works to a better undestanding of epidemiological risk., Dûe à un protozoaire parasitant diverses espèces de mammifères, la cryptosporidiose est une parasitose d’apparition récente en pathologie humaine mais qui a provoqué d’importantes épidémies dans différents pays. Habituellement asymptomatique chez les patients immunocompétents, elle est au contraire généralement très grave chez les immunodéprimés et en particulier chez les sidéens ou chez les transplantés, avec des formes intestinales pures qui peuvent se compliquer de localisations respiratoires. De diagnostic parfois difficile, la cryptosporidiose n’a pas de traitement spécifique réellement efficace. Sa prophylaxie est particulièrement difficile, les formes infestantes (oocystes) généralement véhiculées par l’eau de boisson résistant aux procédés habituels de désinfection industrielle. La mise au point de nouveaux procédés de purification et de tests permettant d’en vérifier les résultats est donc une urgence de santé publique. Sur un plan plus général, on ignore quels ont été les mécanismes de son passage de l’animal à l’homme. Certaines souches se sont d’ailleurs complètement adaptées à ce nouvel hôte et sont génétiquement très différentes des souches d’origine animale. Elles sont responsables de transmissions interhumaines et jouent un rôle probablement essentiel dans les contaminations nosocomiales., Ambroise-Thomas Pierre, Pinel Claudine, Grillot Renée. La cryptosporidiose humaine : une parasitose émergente d’importance croissante en santé publique. In: Bulletin de l'Académie Vétérinaire de France tome 152 n°3, 1999. pp. 91-99.

Anti-Aspergillus fumigatus alkaline protease IgG was investigated by Western blot. Specific IgG antibodies were detected in sera of two of nine patients with proven or highly probable invasive aspergillosis and in sera of one of eight patients with proven or highly probable allergic bronchopulmonary aspergillosis. No response was obtained in sera of 23 control patients. The specific but transient IgG response to this induced enzyme does not recommend its use as sole serodiagnostic aid.

Six different serological methods were tested for their ability to diagnose human hydatidosis. These were indirect immunofluorescence, indirect hemagglutination, counter immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA). In all, 298 sera were examined: 151 sera from negative control subjects, 78 sera from patients with proven hydatidosis, and 69 sera from patients with disease not due to hydatidosis. The last group of samples were used to define the technique's specificity. The specificity and sensitivity of the two recent techniques: enzyme-linked immunoelectrodiffusion assay (ELIEDA) and immunoblotting (electrophoretic migration under denatured and reducing conditions of hydatid fluid antigens, blot and immunoenzymatic assay) were tested mainly on doubtful or weakly positive sera from patients with proven hydatidosis and with filariasis. The combination of enzymelinked immunosorbent assay, indirect immunofluorescence and counter immunoelectrophoresis or ELISA and ELIEDA gave the most dependable and reliable serodiagnostic results with our antigens. ELIEDA is the most useful technique if only one can be used. Our preliminary results with ELIEDA confirm the good specificity and sensitivity of this technique and allow, in some cases, an interpretation of false positive values obtained with ELISA and indirect immunofluorescence. This method also confirms, in most cases, the disease by revealing the arc 5 in patients with low specific IgG content. Immunoblotting in our experimental conditions has a lower sensitivity than ELIEDA but has a specificity for certain proteins (10–12, 15–18, 220, 230 kDa).