Outbreak of cryptosporidiosis in a child day-care centre in Gipuzkoa, Spain, October to December 2011.

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.

A distinctive cytologic pattern for diagnosing tuberculous lymphadenitis in AIDS.

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.

Detection of mycobacterium tuberculosis in paraffin-embedded pleural biopsy specimens by commercial ribosomal RNA and DNA amplification kits.

STUDY OBJECTIVES: To evaluate the utility of two gene amplification systems in historical paraffin-embedded pleural biopsy (PEB) tissues from patients with pleural tuberculosis, and to compare the results to those obtained with conventional histologic and microbiological methods. DESIGN: A retrospective study. PATIENTS AND METHODS: Seventy-four formalin-fixed PEB tissues collected and stored over 12 years (1984 through 1995) were retrieved. Gene amplifications were performed in 57 tissues from patients with diagnoses of pleural tuberculosis and in 17 from patients with carcinoma as controls, using the first version of the Amplified Mycobacterium tuberculosis Direct Test (AMTDT; Gen-Probe; San Diego, CA) and the LCx Mycobacterium tuberculosis Assay (LCxMTB; Abbott Laboratories; Abbott Park, IL). RESULTS: The sensitivities of the AMTDT and LCxMTB were 52.6% and 63.2%, respectively (p = not statistically significant). The specificity of both tests was 100%. Twenty tissue samples (35.1%) were positive by both systems, and 10 tissues (17.5%) were positive only by the AMTDT, while 16 tissues (28.1%) were positive only by the LCxMTB. Both tests gave negative results for 11 specimens (19.3%). When both tests were used, a positive diagnosis was achieved in 80.7% of the samples. Diagnosis of 73.7% of patient conditions had previously been made by smear examination of pleural biopsy and sputum, pleural liquid, or biopsy culture. The overall diagnostic yield with both culture and amplification techniques was 96.5% (55 of 57 patients) for pleural tuberculosis, with amplification techniques adding 22.8% of the diagnoses. CONCLUSIONS: Amplification techniques are useful in archival PEB tissues, providing additional diagnoses beyond culturing, although the sensitivity should be improved, possibly by standardizing protocols.

Evaluation of two different cell lysis methods for releasing mycobacterial nucleic acids in the INNO-LiPA mycobacteria test.

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.

Rapid detection of Mycobacterium tuberculosis in respiratory specimens, blood and other non-respiratory specimens by amplification of rRNA.

SETTING: Diagnostic methods employing gene technology based on amplification of DNA or RNA are expected to improve the speed, sensitivity, and specificity of Mycobacterium tuberculosis detection. The Amplified Mycobacterium Tuberculosis Direct Test (AMTDT) enables the amplification and detection of M. tuberculosis rRNA directly from respiratory specimens. OBJECTIVE: To evaluate the performance of the AMTDT in direct detection of M. tuberculosis in respiratory specimens, blood and other clinical samples, and to compare this method with conventional culture and staining techniques. DESIGN: A total of 554 samples from 450 patients were examined in this study. All clinical specimens (with the exception of bone marrow aspirates and blood samples) were digested and decontaminated with sodium dodecyl (lauryl) sulfate (SDS)-NaOH. Bone marrow aspirates and blood samples were treated with 10% SDS. All processed samples were stained by auramine-rhodamine fluorochrome and inoculated onto Löwenstein-Jensen and Coletsos solid media, and into BACTEC-12B medium. In addition, the blood samples were inoculated into BACTEC 13A medium. The AMTDT was performed according to manufacturer's instructions. In those cases where discrepant results were obtained for AMTDT and cultures, patients' clinical data and other microbiological results were evaluated. RESULTS: The sensitivity, specificity, and positive and negative predictive values for AMTDT were 87.5, 100, 100, and 96.7%, respectively, in respiratory specimens and 86.8, 100, 100, and 92.8%, respectively, in non-respiratory specimens. The differences in sensitivity of these two groups of specimens were not highly statistically significant (P > 0.005). CONCLUSION: The sensitivity and specificity of the AMTDT were satisfactory for detection of M. tuberculosis in all types of clinical samples. Some minor changes in assay format and laboratory protocols may increase the sensitivity of the AMTDT without adversely affecting its specificity.

[Comparative study of acute and chronic forms of pleural tuberculosis]. / Estudio comparativo entre formas agudas y crónicas de tuberculosis pleural.

To analyze whether a characteristic pattern distinguishes acute forms (symptoms starting no more than one week earlier) and subacute or chronic forms (symptoms lasting more than one week) of pleural tuberculosis (PT) in our practice. Retrospective analysis of 102 cases of PT diagnosed between 1986 and 1992, comparing the findings of case histories, imaging, pleural biochemistry and cytology, microbiology of sputum, pleural biopsy and fluid, anatomy and pathology studies of the biopsy specimens, course of disease, response to treatment and sequelae after one year and a half. Thirty-two patients (31.4%) had acute PT and 70 (68.6%) had chronic forms. LDH levels and the percentage of pleural nuclear polymorphism were higher in acute cases, while the concurrence of systemic involvement and lymphocytic predominance was more often seen in chronic cases. Other data analyzed were similar in both groups. No specific clinical or X-ray profiles or differences in course of disease and response to treatment distinguishes between acute and chronic forms of PT after 7 days. Only LDH levels and cytology results differentiated acute forms and, occasionally, such cases were indistinguishable from pneumonia-like effusion.

[Analysis of 90 cases of pleural tuberculosis in relation to adenosine deaminase levels]. / Análisis de 90 casos de tuberculosis pleural en relación a los valores de la adenosina desaminasa.

BACKGROUND: To analyze the adenosine deaminase (ADA) pleural levels and the possibility of a typical pattern of tuberculous pleuritis (TP) in relation to these values. PATIENTS AND METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

[The impact of bronchial colonization in the quality of life of patients with chronic, stable bronchitis]. / Impacto de la colonización bronquial en la calidad de vida del paciente con bronquitis crónica estable.

BACKGROUND: 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). MATERIALS AND METHODS: 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). RESULTS: 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. CONCLUSIONS: 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.

[Drug resistance in Mycobacterium tuberculosis. A multicenter study of the Barcelona area. Grupo de Trabajo sobre Resistencias en Tuberculosis]. / Farmacorresistencia de Mycobacterium tuberculosis. Estudio multicéntrico en el area de Barcelona. Grupo de Trabajo sobre Resistencias en Tuberculosis.

BACKGROUND: 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. PATIENTS AND METHODS: 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. RESULTS: 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. CONCLUSIONS: 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 usefulness of the bone marrow examination in the etiological diagnosis of prolonged fever in patients with HIV infection]. / Utilidad del examen de la médula ósea en el diagnóstico etiológico de la fiebre prolongada en los pacientes con infección por el virus de la inmunodeficiencia humana.

BACKGROUND: 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. PATIENTS AND METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

[Factors associated with pleural thickening in pleural tuberculosis. Review of 99 cases]. / Factores asociados al engrosamiento pleural en la pleuritis tuberculosa. Análisis de 99 casos.

OBJECTIVE: To analyze which factors were related to the development of pleural thickening in pleural tuberculosis (PT). METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

[Cutaneous lymphatic sporotrichosis]. / Esporotricosis cutáneo linfática.

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.

[Complications in Mediterranean boutonneuse fever: prospective study of 78 patients]. / Complicaciones en la fiebre botonosa mediterránea: estudio prospectivo de 78 pacientes.

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.

Risk stratification for dissemination of tuberculosis in HIV-infected patients.

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)

[Pleural tuberculosis: analysis of 105 cases]. / Tuberculosis pleural: análisis de 105 casos.

BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

[Predictive value of neopterin in boutonneuse fever]. / Valor predictivo de la neopterina en la fiebre botonosa.

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.

Comparison of the sodium dodecyl sulfate-sodium hydroxide specimen processing method with the C18-carboxypropylbetaine specimen processing method using the MB/BacT liquid culture system.

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% (P>0.05), respectively, while the specificities were identical (99.7%). The sensitivity of culture was 84.2% and 96.1% (P<0.05), respectively. The time to detection in the MB/BacT liquid culture system was 13.2+/-5.6 and 15.0+/-8.8 days (P>0.05), respectively, and 20.0+/-7.6 and 15.7+/-8.9 days (P<0.05), 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.