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BACKGROUND AND OBJECTIVE: To analyze the characteristics of bone and joint tuberculosis in a Spanish university hospital. PATIENTS AND METHOD: Retrospective study (1984-2006) in a university hospital. All patients had microbiologically and/or histologically proved osteoarticular tuberculosis. Patients with clinical and radiological criteriae, positive tuberculin test responding to antituberculous treatment were not excluded despite negative cultures. RESULTS: We attended 53 patients with osteoarticular tuberculosis (35 males/18 females; mean age: 52 years). Bone tuberculosis involved axial skeleton in 37 patients (71%), peripheral distribution in 12 (21%) and both locations in 4 (8%). Mean time to diagnosis was 8 months. Most common involved joints were knee and ankle. An extraarticular involvement was found in 12 patients (22%) and multifocal bone infection in 6 (11%). Risk factors were present in 22 patients (42%), and 15% were immigrants. The diagnosis was established by a positive culture in 40 cases (75%), and 33 (62%) had suggestive histology. Complications of tuberculosis included medullar compression (9.4%), abscess (12%) and fistulae (9.4%). Spine involvement required surgical intervention in 27% and peripheral involvement in 56%. Curation was achieved in 33 patients (62.3%), curation with secuelae in 19 cases (35.9%) and no consolidation of arthrodesis in one case. CONCLUSIONS: Bone and joint tuberculosis is still common in our area and should be particularly considered in immigrants. Diagnosis delay is remarkable. Complications are present in one third of patients. Surgical treatment is often required.

The INNO-LiPA Mycobacteria Test (Innogenetics, N.V., Belgium) is a PCR-based reverse hybridization assay for the simultaneous identification of several mycobacterial species. We evaluated two simplified lysis methods for mycobacterial DNA release for application in the INNO-LiPA Mycobacteria Test. The two methods were based on either (i) heat treatment or (ii) sonication. Both methods were performed directly on 45 positive liquid cultures (MB-BacT, BioMérieux, Marcy l'Etoile, France) containing 17 different mycobacterial species. These two simple lysis procedures demonstrated similar effectiveness (100%) to that recommended by the manufacturer. They also significantly shortened the time required for mycobacterial DNA release.

The ability of physicians to diagnose tuberculosis is impacted by the use of smear and culture techniques combined with specimen processing methods. The objective of this study was to evaluate the effects of specimen processing on smear and culture sensitivity by comparing the specimen processing method that uses C(18)-carboxypropylbetaine with the method that combines sodium dodecyl sulfate and sodium hydroxide. A total of 1,201 specimens were entered into this study. Specimens were split approximately equally such that one-half of each specimen was processed with sodium dodecyl sulfate-sodium hydroxide, while the other half was processed with C(18)-carboxypropylbetaine. All sediments were subjected to acid-fast staining and then analyzed using the MB/BacT liquid culture system (bioMérieux, France) and solid media. The sensitivity of smear following processing with sodium dodecyl sulfate-sodium hydroxide and C(18)-carboxypropylbetaine was 61.2% and 58.6% (P0.05), respectively, while the specificities were identical (99.7%). The sensitivity of culture was 84.2% and 96.1% (P0.05), respectively. The time to detection in the MB/BacT liquid culture system was 13.2+/-5.6 and 15.0+/-8.8 days (P0.05), respectively, and 20.0+/-7.6 and 15.7+/-8.9 days (P0.05), respectively, on solid media. The contamination rates in the MB/BacT system were 0.8% and 8.7%, respectively, whereas the contamination rates on solid media were 2.6% and 4.3%, respectively. C(18)-carboxypropylbetaine specimen processing was less labor-intensive than sodium dodecyl sulfate-sodium hydroxide processing and improved the ability of laboratory staff to detect the presence of mycobacteria by culture.

Antigen fingerprinting based on surface glycolipid antigens was applied to the epidemiology of clinical isolates of the Mycobacterium avium complex from 128 acquired immunodeficiency syndrome (AIDS) and 31 non-AIDS patients from several different regions of Spain. The application of thin-layer chromatography, gas chromatography-mass spectrometry and monoclonal antibodies, combined with ELISA, allowed a facile identification, differentiation and classification of the isolates. The cumulative results demonstrate that, among the clinical isolates, serovar 4 was predominant in both AIDS (33.6%) and non-AIDS (22.6%) isolates. In general, the results demonstrate geographical as well as disease-related differences in the distribution of Myco. avium complex serovars of clinical importance.

The MB/BacT (Organon-Teknika, USA) is a fully automated, rapid, nonradiometric system for the culture of mycobacteria from clinical samples. The rate of recovery of mycobacteria and the time to detection obtained with the MB/ BacT were compared with those obtained with Löwenstein-Jensen and Coletsos solid media and Bactec 7H12 (12B) (Becton-Dickinson, USA) broth when 600 processed specimens were inoculated into all media in parallel. Specimens included 383 respiratory samples, 20 urine samples, 23 purulent exudates, 13 stool samples, 103 blood samples, 14 bone marrow aspirates, and 44 body fluid samples or aspirates. Overall, 106 mycobacterial isolates comprising six species were recovered, of which 100 (94.3%) were detected with MB/BacT, 98 (92.5%) with egg-based media, and 96 (90.2%) with Bactec 12B. The recovery rates of Mycobacterium tuberculosis complex with MB/BacT, egg-based media, and Bactec 12B were 98.7%, 93.7, and 89.9%, respectively. The average number of days to detection of single mycobacterial isolates was 14.2 days for MB/BacT, 26.1 days for egg-based media, and 11.7 days for Bactec 12B. The contamination rates were higher in MB/BacT (5%) than in Bactec 12B (1.8%) or egg-based media (1.5%). MB/BacT is a reliable, nonradiometric, less labor-intensive alternative to Bactec 12B for recovery of mycobacteria, but, as with other liquid culture methods, MB/BacT should be used in combination with a solid medium, not on its own.

We evaluated a newly commercial enzyme immunoassay (EIA) (Biotest Legionella Urin Antigen EIA; Biotest AG, Dreieich, Germany) for detection of antigens of all Legionella pneumophila serogroups with a relatively wide spectrum of cross-reactivity as well as antigens of other Legionella spp. by comparing its sensitivity and specificity with those of an EIA for detection of L. pneumophila serogroup 1 antigen ( Legionella urinary antigen EIA; Binax, Portland, Maine). Both tests were performed with both concentrated and nonconcentrated urine samples. We also evaluated the capabilities of both EIAs to detect extracted soluble antigens of American Type Culture Collection (ATCC) Legionella strains ( L. pneumophila serogroups 1 to 14, L. bozemanii , and L. longbeachae ). The sensitivity of the Biotest EIA was 66.66% in nonconcentrated urine and 86.66% in concentrated urine. The sensitivity of the Binax EIA was 63.76% and 88.88% in nonconcentrated and concentrated urine, respectively. The specificity was 100% in nonconcentrated and concentrated urine for both assays. The Binax EIA and Biotest EIA detected extracted soluble antigens of L. pneumophila serogroups 1 to 14 and L. bozemanii ATCC strains. The cross-reactions observed with the Binax EIA were probably due to common epitopes directly related to lipopolysaccharide. Further studies are required to determine the usefulness of the Binax EIA for detection of urinary antigens from Legionella species and serogroups other than L. pneumophila serogroup 1. The Biotest EIA proved to be as rapid, sensitive, and specific as the Binax EIA for the diagnosis of legionellosis. Concentration of antigen present in urine increased the sensitivities of both techniques with no reduction in specificity.

El objetivo de este estudio ha sido analizar la posibilidad de un patron caracteristico que distinga entre formas agudas (sintomatologia inicial igual o inferior a una semana) y subagudas o cronicas (superior a una semana) de tuberculosis pleural (TP) en nuestro medio. Hemos llevado a cabo un analisis retrospectivo de 102 TP diagnosticadas entre 1986 y 1992, comparando los hallazgos de la historia clinica, radiologia, bioquimica y citologia pleural, microbiologia del esputo, liquido y biopsia pleural, estudio anatomopatologico de las biopsias, evolucion, respuesta al tratamiento y secuelas al ano y medio de control. Del total de casos, 32 eran de TP aguda (31,4%) y 70 no aguda (68,6%). Los datos analizados fueron similares en ambos grupos a excepcion de valores mas elevados de LDH y del porcentaje de polimorfonuclea- res pleurales en las TP agudas, mientras en las no agudas se observo una mayor asociacion de un sindrome constitucional y un predominio linfocitario. En la TP no se confirma un patron clinico o radiologico especifico ni diferencias evolutivas y de respuesta al tratamiento que distingan entre formas agudas o no a partir de los 7 dias. La LDH y la citologia fueron los unicos parametros que diferenciaron las formas mas agudas y, en ocasiones, inicialmente se mostraron indistinguibles de un derrame paraneumonico.

Setting: The resurgence of tuberculosis and the increase of Mycobacterium avium complex infections have renewed interest in developing more efficient systems for isolation of mycobacteria. The Bactec radiometric method enhances the recovery and shortens the detection time of mycobacteria; however, many clinical laboratories are subject to severe restrictions on the utilization of 14C radiolabelled culture media. Objective: To compare the biphasic MB-Check system in the recovery of mycobacteria from clinical specimens with the Bactec-460 system and Lowenstein-Jensen (LJ) medium. Design: The MB-Check was evaluated for routine use in a clinical laboratory by comparing the rates and times for isolation of mycobacteria from 1840 clinical specimens with results obtained using the other systems. The isolated organisms were identified by standard procedures and specific deoxyribonucleic acid (DNA) probes. Results: The rate of recovery of all mycobacteria with the MB-Check system was 84.8%, compared to 87.5% for the Bactec-460 system and only 64.2% on LJ. Of the 147 M. tuberculosis isolates recovered by all methods combined, 92.5 % were recovered by MB-Check, 87.1% by Bactec and 79.6% on LJ. Of the 109 isolates of nontuberculous mycobacteria (NTM) recovered by all methods combined, 44% were recovered on LJ, 75% by MB-Check and 88% by Bactec (P < 0.001). A combination of the Bactec and MB-Check systems allowed recovery of 99.2% of isolates. The Bactec system showed a faster detection time than the other two methods. Conclusion: The MB-Check and Bactec systems were more efficient than LJ medium for the isolation of both M. tuberculosis and NTM. The performance of the MB-Check system and the ease with which processing, inspection, detection, and isolation can be performed, indicated that the biphasic approach for cultivation of mycobacteria is feasible and practical. The MB-Check system is more suitable than Bactec system for laboratories that have a small number of specimens or in situations where the disposal of radioactive waste is a problem.

The lower airways of asymptomatic chronic obstructive pulmonary disease (COPD) patients can be colonized by bacteria, mainly Haemophilus influenza, Streptococcus pneumoniae, and Moraxella catarrhalis. However, the role of lower airway bacteria in stable and exacerbated COPD has not been well defined. To determine the importance of lower airway bacterial infection in COPD we studied 40 outpatients with stable COPD (Group A: age 61.1 +/- 9.9 yr; [mean +/- SD]; FEV1/FVC 51.7 +/- 12.5) and 29 outpatients with exacerbated COPD (Group B: age 63.4, SD 9.0 yr; FEV1/FVC 52.0, SD 9.6), using the protected specimen brush (PSB) for microbiology sampling. Group A consisted of outpatients with stable COPD having normal or near-normal chest X-rays, with clinical indications for performing fiber-bronchoscopy (pulmonary nodule, remote hemoptysis); Group B consisted of patients with exacerbated COPD who voluntarily accepted lower airway microbiology sampling. To avoid contamination by upper airway flora the PSB was used for bacterial sampling in all the cases and concentrations > or = 1,000 colony-forming units/milliliter (CFU/ml) were considered positive. Results were as follows: Group A: Lung function data in outpatients with stable COPD were lower than the reference values for this population (FVC 2.97 +/- 1.02 L, FVC% 71.4 +/- 22.4, FEV1 1.59 +/- 0.79 L, FEV1% 51.2 +/- 23.0). Positive PSB cultures were obtained in 10 of 40 cases (25%), mainly of H. influenzae and S. pneumoniae. Two of 40 cases had positive cultures at concentrations > or = 10,000 CFU/ml (5.0%).(ABSTRACT TRUNCATED AT 250 WORDS)

We evaluated a commercially available radioimmunoassay technique for detecting the soluble antigen of Legionella pneumophila serogroup 1 using concentrated and nonconcentrated urine. Concentration was achieved by selective ultrafiltration. The sensitivity of the technique was 60.9% for nonconcentrated urine and 80.4% for concentrated samples, with 100% specificity in both cases.

A detailed study comparing specimen processing methods for the isolation of mycobacteria was recently reported [1]. In the follow-up study presented here, 1,201 specimens (95% respiratory) were collected and split approximately equally such that one-half of each specimen was processed with 3% sodium dodecyl sulfate-1% sodium hydroxide (SDS-NaOH) [2] and the other half was processed with C18-carboxypropylbetaine (CB-18) followed by lytic enzyme decontamination [3, 4]. All processed specimens were analyzed by culture using the MB/BacT liquid culture system containing the antibiotic supplement MAS (bioM rieux, France). The SDS-NaOHprocessed specimens were also inoculated onto L wenstein-Jensen slants, while the CB-18-processed specimens were also inoculated onto 7H11-selective plates (50 g/ml carbenicillin, 200 U/ml polymyxin B, 10 g/ml amphotericin B, and 20 g/ml trimethoprim lactate). CB-18/lytic enzyme processing increased overall culture sensitivity by approximately 14.1% (P

The ability of a polymerase-chain-reaction-based technique combined with a reverse hybridisation line-probe assay to detect and identify eight of the most clinically significant mycobacterial species directly from cultures in liquid medium was evaluated. The line-probe assay allows simultaneous identification of eight different mycobacterial species. Ninety-seven mycobacterial strains belonging to 22 different species were evaluated. All strains were previously inoculated into MB/BacT bottles (Organon Teknika, USA). The sensitivity and specificity of the test when applied on positive MB/BacT liquid cultures containing isolates from mycobacterial species included as specific probes on the line-probe assay strip was 100% and 100%, respectively. These results further support the potential clinical usefulness of this technique in the diagnosis of mycobacterial infections.

The aims of this multicenter study was to establish the level of primary and acquired drug resistance of M. Tuberculosis strains isolated in Barcelona and to identify possible risk groups using clinical data.All tuberculosis patients with isolation and identification of M. tuberculosis strains from October 1995 to September 1997 were included. Susceptibility tests isoniazid, rifampin, ethambutol, streptomycin and pyrazinamide were performed using the Bactec 460 system and the proportions method on solid medium. Logistic progression was used for statistical analysis.The total number of patients included was 1,749 (1,535 non-treated and 214 previously treated). Primary drug resistance was 5.7% (isoniazid 3.8%; rifampin 1.0%, streptomycin 2.1%, ethambutol 0.3% and pyrazinamide 1.0%). Acquired drug resistance was 20.5% (isoniazid 17.3%, rifampin 9.8%, ethambutol 1.9%, streptomycin 4.7% and pyrazinamide 6.5%). Primary drug resistance was associated with people over 60 years old and women.The low level of drug resistance enables antituberculosis treatment of non-treated patients to start with the standardised three-drug regimes except in the case of foreign people from countries with a high level of drug resistance. Susceptibility tests are recommended on all M. tuberculosis strains isolated, together with controlled studies of drug resistance surveillance.

The performance was evaluated of a fluorescence in situ hybridisation assay using peptide nucleic acid probes (Dako Probe MTB Culture Confirmation Test; Dako, Denmark) for identification of Mycobacterium tuberculosis complex (MTC) organisms and differentiation between tuberculous and non-tuberculous mycobacteria (NTM) in material taken directly from Bactec 12B (Becton Dickinson, USA) and MB/BacT (Organon Teknika, USA) bottles. The test was applied to 129 smear-positive (Ziehl-Neelsen stain) clinical specimens, 48 previously identified clinical strains of mycobacteria (12 MTC and 36 NTM), and 51 reference strains (7 MTC and 44 NTM) which were all previously inoculated into Bactec 12B and MB/BacT bottles. The sensitivity and specificity of the assay for MTC-positive cultures was 87.6% and 100%, respectively, for Bactec 12B, and 100%, respectively, for MB/BacT. The sensitivity and specificity of the assay for NTM-positive cultures was 100% for both media.

The aim of this study was to prospectively analyze the bacterial etiology of community-acquired pneumonia in adults in Spain. From May 1994 to February 1996, 392 episodes of CAP diagnosed in the emergency department of a 600-bed university hospital were studied. An etiological diagnosis based on noninvasive microbiological investigations was achieved in 228 cases (58%); 173 of these diagnoses were definitive and 55 probable. Streptococcus pneumoniae, which caused 23.9% of the episodes, was the predominant pathogen observed, followed by Chlamydia pneumoniae (13.5%) and Legionella pneumophila (12.5%). Other less frequent pathogens found were Haemophilus influenzae (2.3%), Pseudomonas aeruginosa (1.5%), Mycoplasma pneumoniae (1.3%), Coxiella burnetii (1%), Moraxella catarrhalis (2 cases), Nocardia spp. (2 cases), and Staphylococcus aureus (2 cases). Streptococcus pneumoniae was significantly more frequent in patients with underlying disease and/or age > or =60 years (28% vs. 13%, P = 0.002), while Legionella pneumophila was more frequent in patients below 60 years of age and without underlying disease (20% vs. 9%, P = 0.006). Likewise, Streptococcus pneumoniae and Legionella pneumophila were the most frequent etiologies in patients requiring admission to the intensive care unit, occurring in 29% and 26.3% of the patients, respectively. In addition to Streptococcus pneumoniae, other microorganisms such as Chlamydia pneumoniae and Legionella spp. should be seriously considered in adults with community-acquired pneumonia when initiating empiric treatment or ordering rapid diagnostic tests.

To analyze the usefulness of bone marrow (BM) study in the diagnosis of fever of unknown origin (FUO) in patients infected by the human immunodeficiency virus (HIV) in a single center during a period of 42 months.182 episodes of FUO in 162 patients p3th HIV infection were studied. Age, sex, risk factor for HIV infection, hemoglobin level, counts of leucocytes, neutrophils, lymphocytes, CD4 positive lymphocytes, platelets and levels of hepatic enzymes, albumin and beta 2-microglobulin were studied. BM aspirate was performed in all episodes for cytologic and microbiologic examination, and BM biopsy was done in 43. Analysis of factors related with the probability of diagnosis by BM examination was carried out.The median age was 33 years (range, 22-70), and 123 were males. Drug abuse was the most frequent risk factor for HIV infection (63%). One hundred thirty patients had previous AIDS diagnosis before they were evaluated for unexplained fever. A specific diagnosis was achieved in 161 episodes (88%) and the most frequent diagnoses were Mycobacterium spp. (55%) and Leishmania spp. (14%) infections. Fifty-four episodes (30%) were diagnosed by BM examination, and in 36 (20%) BM study was the only diagnostic tool. Examination of the BM aspirate yielded the diagnosis in 40 out of the 178 episodes (13%), whereas BM biopsy was a diagnostic tool in 8 (19%); in 9 additional cases (21%) granulomas were observed. Microbiologic study of BM smears for mycobacterial infections was positive in 28 of the 143 episodes (19%), and the culture for Leishmania was positive in 2 out of the 42 cases. The presence of thrombocytopenia (75 x 10(9)/l) and elevated serum levels of aspartate-aminotransferase (AST) (100 U/l) were the factors associated with a high probability to obtain the diagnosis through BM study.In patients infected by the HIV and unexplained fever, BM examination is an useful procedure for the diagnosis, particularly in areas where infections by Mycobacterium spp. and Leishmania are prevalent. So that, in our setting, systematic use of this procedure is justified for diagnosis of FUO in those patients.

The aim of this study was to determine the impact of respiratory function and bacterial colonization of the lower airway on the quality of life of patients with chronic, stable bronchitis (CB).A series of 41 patients with stable CB was studied (age: 63.8; standard deviation (SD) 9.1 years; FVC% 91.0 (18.9); FEV1% 74.6 (23.7); FEV1/FEC 62.8 (11.2) with normal thoracic radiography. Patients with previous diagnosis of bronchiectasia, bronchial asthma and/or positive bronchodilatory tests (15%) were not included in the study. Bacterial growth in a sputum sample of grade 4-5 of the Murray-Washington scale was considered diagnostic of bronchial colonization. Measurement of the quality of life was performed with the Nottingham Health Profile (NHP) and the St. George Respiratory Questionnaire (SGRQ).The patients presented a moderate alteration in their quality of life with scores over 25 in most of the dimensions of the NHP and the SGRQ. In 9 out of 41 cases (22%), the sputum cultures demonstrated bronchial colonization with the most frequently isolated bacterias being Haemophilus influenzae and Moraxella catarrhalis. Multivariate analysis performed with the quality of life as the dependent variable showed an association between FEV1/FEC1 and the SGRQ score (R2 = 0.18), and energy (R2 = 0.09) and physical mobility (R2 = 0.05) of NHP.Bronchial obstruction is the main determinant in the quality of life in patients with stable CB. Colonization of the lower airway is observed in 22% of the patients and also influences the quality of life of the patients but to a much lesser extent.

Two commercial systems for the amplification and detection of Mycobacterium tuberculosis directly from respiratory samples were compared. The Roche Cobas Amplicor MTB Test and the Roche manual Amplicor MTB Test (Roche Diagnostic Systems, USA) were applied to 755 decontaminated respiratory specimens collected from 470 patients. Results were compared with those of acid-fast staining and culture. A total of 251 specimens were collected from 156 patients diagnosed with pulmonary tuberculosis, including 28 specimens corresponding to 13 patients that were receiving antituberculous treatment. Given the overall positivity rate of 33.2% (251/755), the sensitivity, specificity, and positive and negative predictive values were 92.4, 100, 100, and 96.5%, respectively, for the Cobas Amplicor MTB Test and 90.8, 100, 100, and 95.8%, respectively, for the Amplicor MTB Test. For 204 (81.3%) smear positive specimens and 47 (19.7%) smear negative specimens, the sensitivity values were 100 and 59.6%, respectively, for the Cobas Amplicor MTB Test and 100 and 51%, respectively, for the Amplicor MTB Test. There were no statistically significant differences in sensitivity or specificity between the two assays and culture (p0.05). The overall results of both assays were concordant for 99.5% of the samples. It is concluded that although both nucleic acid amplification methods are rapid and specific for the detection of Mycobacterium tuberculosis complex in respiratory specimens, the Cobas Amplicor MTB Test appears to be slightly more sensitive than the Amplicor MTB Test when smear negative specimens are investigated.

The clinical course of legionella pneumonia in immunosuppressed patients is uncertain. This study was undertaken to determine the clinical evolution of legionellosis on the basis of the immune state and to establish the variables associated with death directly related to legionellosis. The study included 78 patients: 28 with chronic disease who had received immunosuppressive treatment (group 1), 24 with chronic disease without immunosuppressive treatment (group 2), and 26 controls. Inclusion criteria were the occurrence of nosocomially acquired pneumonia, Legionella pneumophila infection, and erythromycin therapy that was initiated within 72 hours following diagnosis. Respiratory and extrarespiratory complications were observed more frequently in groups 1 and 2. Bilateral radiological involvement was most frequent in group 1, and recurrence of legionella pneumonia was observed exclusively in group 1. None of these variables achieved statistical significance. The global mortality of the series was 11.5% (17.9%, 12.5%, and 3.8% in groups 1, 2, and 3, respectively). Variables statistically related to mortality were acute renal failure, shock, and need for mechanical ventilation. Although many of the variables analyzed lacked statistical significance, a trend was seen between complications and basal immunosuppression, as previously suggested.

A sodium dodecyl (lauryl) sulfate method was evaluated for the preparation of blood specimens and bone marrow aspirates for use in two amplification procedures (Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test [AMTDT] and Roche Amplicor M. avium/M. intracellulare [MAI] Test) for the detection and identification of Mycobacterium tuberculosis and M. avium and M. intracellulare, respectively. The AMTDT is based on amplification of rRNA, whereas the Amplicor MAI Test amplifies a specific DNA region of the 16S rRNA gene. The results of amplification techniques were compared with those of standard culture and culture in BACTEC 13A and BACTEC 12B liquid media. A total of 121 blood specimens and 15 bone marrow aspirates were collected from 136 AIDS patients. Mycobacterial growth was recovered for 103 specimens; 35 yielded M. tuberculosis, 62 yielded M. avium, 5 yielded M. genavense, and 1 yielded M. kansasii. The values of sensitivity and specificity in pretreated specimens for detection of M. tuberculosis by the AMTDT were 94.3 and 100%, respectively, and those for detection of M. avium by the Amplicor MAI Test were 91.9 and 100%, respectively. The simple lysis method described in the present work allows the recovery of mycobacteria from blood specimens and bone marrow aspirates and may be used in combination with the AMTDT and the Amplicor MAI Test to detect and identify different members of the genus Mycobacterium. This method might also be applicable for the identification of mycobacteria from blood culture fluids with acridinium-ester-labeled DNA probes.

To analyze the adenosine deaminase (ADA) pleural levels and the possibility of a typical pattern of tuberculous pleuritis (TP) in relation to these values.We analyzed 90 consecutive cases of TP dividend into two groups on the basis of the pleural ADA measurements: group 1 (less than 43 U/I) and group 2 (equal or higher than 43 U/I). We compared: age, sex, tuberculous risk factors, clinic onset and duration, PPD, chest roentgenogram, biochemical (LDH, glucose and proteins) and cytological examination of the pleural fluid, microbiologic studies on sputum, pleural fluid and biopsies, histologic findings of the biopsies, evolution and sequelae after the treatment.36 patients in the group 1 (40%) and 54 in the group 2 (60%) (p = 0.01). All the parameters were similar in both groups excluding LDH pleural levels, that were higher in group 2. In 20 cases of the group 1, we obtained a second analysis of pleural ADA and we observed in 11 cases (55%) an ADA value above 43 U/I.We confirmed that ADA pleural level is increased in the majority of our patients with a low initial sensibility of 60%, that can increase with a second determination (72%). ADA activity below 43 U/I cannot exclude the tuberculous aetiology of an effusion and can persist negative in other analyses. The results suggest a response of different chronology without any relation to initial ADA pleural level.

To analyze which factors were related to the development of pleural thickening in pleural tuberculosis (PT).We reported 99 patients diagnosed as having PT separated into two different groups according to the presence of radiographic pleural thickening after completing the treatment: normal chest roentgenogram (group 1) or pleural thickening (group 2). We compared: clinical history, chest radiography, pleural fluid analysis, microbiological studies and effects of the treatment.35 cases in group 1 (35.35%) and 64 in group 2 (64.45%). All the compared results did not differ between the two groups, excluding a higher incidence of febrile cases, increased white cell count and decreased relative percentage of pleural lymphocytes in group 1, and increased number of cases with hemoptysis, positive sputum cultures in patients with parenchymal infiltrates, relative percentage of pleural lymphocytes and decreased white cell count in group 2.We reported an elevated incidence of residual pleural thickening, but it is unlikely that all the cases could have any clinical significance. We didn't confirm a characteristic pattern. Our findings suggested that a different reaction related or not to the hypersensitivity, rather than the inflammatory response to infection was the responsible for the fibrosis.

We present the results of a retrospective study with patients diagnosed of tuberculous pleuritis (TP) to offer a view and a perspective of this entity in our area.We reviewed 105 cases of TP according to age, tuberculous risk factors, clinical onset, tuberculin test, chest roentgenogram, pleural fluid analysis (ADA, LDH, glucose and proteins) and pleural biopsy, and the evolution or sequels of the pleural effusion after a specific treatment.TP was a disease of younger patients (62% less than 30 years old) with tuberculous risk factors (34.3%), positive tuberculin test (81%) and an acute or subacute onset (88.5%). The pleural effusion was unilateral (98.1%), small-sized (61%), with pulmonary infiltrates in the 22.4%. All of the effusions were exudates with the exception of a transudative case. The levels of LDH, glucose and ADA were increased and the white cell counts below 6000/mm2, predominantly mononuclear. ADA was below 40 U/L in the 17% of cases. Pleural biopsy established the diagnosis in the 92.2% and the diagnosis was obtained in the 98.6% when we associated the analysis of ADA, pleural fluid and pleural biopsy. Specific treatment was accompanied with a good response.Our findings supported that TP maintained the predilection for younger patients with tuberculous risk factors. The clinical onset, radiographies, pleural fluid analysis and the utility of all diagnostic examinations were similar to other previous studies with the exception of few atypical cases. We confirmed a good prognosis with a correct treatment.

From January 1988 to October 1992, the primary resistance to first-line antituberculous drugs in 501 tuberculous patients was evaluated prospectively. Three-hundred and seventeen patients were HIV-negative and 184 were HIV-positive; these patients had several different clinical forms of tuberculosis. Moreover, the acquired resistance to antituberculous drugs was studied in 295 non-AIDS patients and in 42 AIDS patients with evidence of antecedent tuberculosis treatment. The data indicated that during these five years there was no consistent and clear-cut trend toward greater frequency of primary drug resistance to any of the first-line antituberculous drugs. Primary drug resistance in HIV-positive patients (7.1 %) did not differ significantly (p>0.05) from that found in HIV-negative patients (8.2 %). Among HIV-positive patients, the acquired drug resistance pattern was similar to that detected in HIV-negative patients although the frequency of resistance in the former (69 %) was significantly higher (p

Tuberculous lymphadenitis (TL) is a very common infection in human immunodeficiency virus (HIV)-infected patients. We performed fine-needle aspiration biopsy (FNAB) of enlarged lymph nodes in 57 HIV-infected patients to evaluate its usefulness in this population. We observed three cytologic patterns in 21 patients diagnosed as having TL: granulomatous lymphadenitis (GL) in 4 FNABs, necrotizing granulomatous lymphadenitis (NGL) in 7 FNABs, and necrotizing lymphadenitis (NL) in 12 FNABs. GL and NGL are already well-known and considered to be highly suggestive of TL. Our results support the idea that NL should have the same diagnostic value as GL or NGL. In the group of 12 patients with NL, TL was confirmed in 11 by microbiologic methods (7 by a positive Ziehl-Neelsen stain and 4 by a positive Löwenstein culture) and in the remaining patient by a biopsy that showed NGL with acid-fast bacilli. We conclude that FNAB is a useful, inexpensive, and safe technique for diagnosing TL in HIV-infected patients. The finding of a NL pattern is suggestive enough of TL to start antituberculous treatment.

The location of tuberculosis (TB) early in the course of HIV-induced immunosuppression was located, and an attempt was made to determine the correlation between the degree of immune suppression and prognostic variables to stratify the risk for dissemination of TB in HIV-infected patients. Clinical and laboratory characteristics were reviewed in 73 HIV-infected patients with TB admitted between 1984 and 1990. The presence of Mycobacterium tuberculosis was investigated in different clinical samples to verify the diagnostic yield of different sources. TB was extrapulmonary in 46.6 per cent of patients in whom it was their first opportunistic infection, and in 46.7 per cent of patients with previously diagnosed AIDS (p = NS). TB was frequently associated with other opportunistic infections, particularly oesophageal candidiasis (p = 0.006). Patients with localized extrapulmonary or disseminated TB presented more often with cytopenias, hypoalbuminaemia and oral thrush. The existence of extrapulmonary TB or another opportunistic coinfection allowed AIDS to be diagnosed in the same admission in 30 patients and a mean of 8.4 months later in another eight. Extrapulmonary TB was found to be as common in early HIV infection as in patients with established AIDS. Haematological derangements were common in these patients, and cytopenias, hypoalbuminaemia and oral thrush were useful predictors of TB dissemination. The location of TB and its dissemination were not significantly linked to a more advanced CDC stage of HIV infection or a more profound fall in CD4 count.(ABSTRACT TRUNCATED AT 250 WORDS)

A new case of lymphocutaneous sporotrichosis is reported. We emphasize the low number of cases reported in the Spanish literature so far. We think that the incidence of this condition is higher than is reported. Because near all published cases had been published by the same authors who are familiarized and possess a high suspicion rate of this condition.

Neopterin is an intermediate purine metabolite which can be determined in serum and has been shown to be elevated in processes that stimulate cell mediated immunity. Neopterin serum levels have been measured in 37 patients diagnosed of boutonneuse mediterranean fever. Serum titers were higher during the acute phase than during the convalescence in 97.2% of patients. Neopterin levels normalized as the disease improved clinically. On the other hand, the highest levels were found in patients with a torpid evolution and appearance of complications.

A prospective study of the clinical evolution of 78 patients diagnosed of mediterranean botonous fever has been performed. Hospital admissions because of complications were necessary in 5 (6,4%) patients. 13 patients (16,6%) had a devious evolution with signs of severe involvement of organs and systems. The evolution was favorable in the remaining 60 cases (79,9%) on home treatment with doxycycline. The unfavorable evolution was associated with advanced age, underlaying chronic disease, and the delay in the initiation of the appropriate treatment. Previous thrombopenia was the major predictive biological factor of unfavourable evolution in our patients.

Five hundred twenty processed respiratory specimens from 326 patients received for the diagnosis of tuberculosis or other mycobacterial infections were tested by means of the LCx Mycobacterium tuberculosis Assay from Abbott Laboratories, which uses ligase chain reaction technology for the direct detection of M. tuberculosis complex in respiratory specimens. The results of the LCx M. tuberculosis Assay were compared with the results of culture and staining techniques. After a combination of culture results and the patient's clinical data, a total of 195 specimens were collected from 110 patients who were positively diagnosed as having pulmonary tuberculosis. Twenty-three of these 195 specimens which corresponded to 10 patients with a history of pulmonary tuberculosis (TB) and anti-TB treatment ranging from 1 to 6 months were culture negative. The other 172 specimens were culture positive for M. tuberculosis. With an overall positivity rate of 37.5% (195 of 520 specimens), the sensitivity, specificity, and positive and negative predictive values were 90.8, 100, 100, and 94.7%, respectively, for the LCx M. tuberculosis Assay; 88.2, 100, 100, and 93.4%, respectively, for culture; and 82.6, 92, 72.9, and 97.6%, respectively, for acid-fast staining. For 161 specimens (82.6%) from patients smear positive for the disease and 34 specimens (17.4%) from patients smear negative for the disease, the sensitivity values for the LCx M. tuberculosis Assay were 98.8 and 53%, respectively. There were no statistically significant differences in the sensitivities and specificities between the LCx M. tuberculosis Assay and culture (P > 0.05). Conclusively, the LCx M. tuberculosis Assay has proved to have an acceptable sensitivity and a high specificity in detecting M. tuberculosis and has the potential of reducing the diagnosis time to an 8-h working day.

The concentrations of sodium dodecyl (lauryl) sulfate (SDS) found in the sediments of the respiratory samples pretreated with SDS-NaOH varied between 3.36 and 12.42 mg/ml. These concentrations of SDS were higher than the level considered critical (> or = 0.16 mg/ml) to obtain negative results with the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test. Extensive washing, after neutralization of sediment, is sufficient to remove all traces of detergent which interfere with the enzymes used by the Mycobacterium Tuberculosis Direct Test assay.

From October to December 2011, an outbreak of 26 cases of cryptosporidiosis occurred in a day-care centre in Gipuzkoa, Spain. The infection spread from person to person and affected 24 children under two years of age (attack rate: 38%) and two caregivers. Cryptosporidium oocysts were observed in 10 of 15 samples. During 2010, only four cases of cryptosporidium were detected in Gipuzkoa, and 27 overall in Spain.

We evaluated a commercial enzyme immunoassay (EIA) kit for detection of Legionella pneumophila serogroup 1 soluble antigen by comparing it to radioimmunoassay (RIA), using both concentrated and nonconcentrated urine samples. The sensitivity of EIA was 67.4% in nonconcentrated urine samples and 82.6% in concentrated urine samples. The sensitivity of RIA was 60.9% and 84.8% in nonconcentrated and concentrated urine samples, respectively. Our study indicates that the sensitivity and specificity of EIA are comparable to those of RIA, and that concentrating the antigen by selective ultrafiltration increases sensitivity for both EIA and RIA, with no significant decrease in specificity.

A commercial assay (Inno-Line Probe Assay; Innogenetics, Belgium) was evaluated to determine its ability to detect rifampin resistance in Mycobacterium tuberculosis directly from clinical specimens. Fifty-nine selected specimens (42 respiratory and 17 nonrespiratory) culture positive for Mycobacterium tuberculosis were tested along with their corresponding isolates in culture. The results were compared with those obtained by in vitro susceptibility testing. The results of the line probe assay to detect rifampin resistance in Mycobacterium tuberculosis present in clinical specimens and in cultured isolates were concordant for 58 of 59 (98.3%) isolates (95% confidence limits = 90.9-99.9%). The line probe assay failed only once, when a fecal specimen was tested; no amplification was observed due to the presence of inhibitory compounds. The most frequently observed mutation was His526--Asp (58.7%), followed by the His526--Tyr (23.9%); together, they represented 82.6% of rifampin-resistant samples. In conclusion, the Inno-Line Probe Assay is a rapid, useful method for detecting the presence of Mycobacterium tuberculosis complex and its resistance to rifampin directly from clinical specimens and culture. Moreover, since rifampin resistance is a potential marker for multidrug resistance in Mycobacterium tuberculosis, this assay may constitute an important tool for the control of tuberculosis.