Identification and Susceptibility Testing on Samples of Mycobacterium spp.

Background: Tuberculosis is still a major global health problem. Human tuberculosis is caused by species of bacteria belonging to the Mycobacterium genus. In this study we determined mycobacterial species affecting patients from Botucatu, Brazil, and tested M. tuberculosis sensitivity to different drugs. Methods: Data were obtained from Clinical Laboratory Analysis records at Botucatu Medical School University Hospital, UNESP. All samples were processed according to standard isolation procedures from the 2008 Brazil Ministry of Health Mycobacteria Manual, which consist of staining smears by the Ziehl-Neelsen technique and seeding cultures in the Löwenstein-Jensen medium. Results: Samples were isolated from sputum (80.5%), bronchoalveolar lavage (13.8%), pleural fluid (4.6%), and cerebrospinal liquor (1.1%). Smears were evaluated in 87 cases and a total of 59 patients showed positive smears; 55 from 70 sputum samples and 4 from 12 bronchoalveolar lavage samples. No pleural fluid (4) or cerebrospinal liquor (1) samples showed positive smears. The most commonly identified strain was M. tuberculosis (61 cases); followed by M. avium and M. gordonae 2 cases each, and M. peregrinum and M. abscessus 1 case each. Mycobacteria were not identified in 20 patients. Only two strains of M. tuberculosis were multidrug resistant; one was resistant to isoniazid, rifampicin, and pyrazinamide. These two patients evolved to cure. Conclusion: This study highlights a small but troubling percentage of multidrug resistant samples and reveals the occurrence of nontuberculous mycobacteria, emphasizing the importance of correctly identifying species and testing sensitivity to antibacilar drugs to assure an adequate therapy.

PCR identification of Mycobacterium tuberculosis complex in a clinical sample from a patient with symptoms of tuberculous spondylodiscitis

A 42-year-old male complaining of thoracic spine pain was admitted to the hospital for evaluation. An X-ray and computer tomography of the thoracic spine showed spondylodiscitis of the L3 lumbar and L2-L3 intervertebral disk. The tuberculin skin test (PPD) was strongly positive. A radioscopy-guided fine needle aspirate of the affected area was cultured but did not reveal the cause of the disease. Two biopsy attempts failed to reveal the cause of the disease by culturing or by acid-fast-resistant staining (Ziehl Neelsen) of the specimens. A third biopsy also failed to detect the infectious agent by using microbiological procedures, but revealed the presence of a 245-bp amplicon characteristic of the Mycobacterium tuberculosis complex after PCR of the sample. The result demonstrates the efficacy of PCR for the identification of M. tuberculosis in situations in which conventional diagnosis by culturing techniques or direct microscopy is unable to detect the microorganism. Following this result the patient was treated with the antituberculous cocktail composed by rifampicin, pirazinamide and isoniazid during a six-month period. At the end of the treatment the dorsalgia symptoms had disappeared.

Levels of C-reactive protein in serum samples from healthy children and adults in Säo Paulo, Brazil

C-reactive protein (CRP) was measured by ELISA in the sera of 165 healthy blood donors and 125 normal children 1 to 14 years old. The serum levels of blood donors ranged from 0.05 to 57.6 mg/l with median and mean values of 1.8 ml/l and 4.86 mg/l, respectively. CRP levels ranged from 0.02 to 14.4 mg/l in the children's sera, the median being 0.45 mg/l and the mean 1.65 mg/l. No individual lacking CRP was detected. The high CRP levels observed in the present study suggest that the population of the State of Sao Paulo may usually be exposed to subacute infections and/or inflammation without presenting clinical symptoms.

Evaluation of diagnostic tests for human leptospirosis

The IgM-PK-ELISA, an enzyme-linked immunosorbent assay for immunoglobulin M employing a proteinase K-treated antigen, and the "Leptoteste-S" macroagglutination test were evaluated for use in a rapid serodiagnosis of human leptospirosis. The microscopic agglutination test (MAT) was used as reference. The three serological tests were applied to serum samples from patients with leptospirosis (N=89), typhoid fever (N=8), malaria (N=19), syphilis (N=20), hepatitis (N=16) and from clinically healthy donors (N=92). The overall results of the IgM-PK-ELISA and the "Leptoteste-S"are comparable to those of the MAT. However, both tests differed statistically from MAT in terms of the positivity of the acute-phase sera, with approximately 38 per cent of the patients with leptospirosis being identified earlier than when MAT was used. The IgM-PK-ELISA, with 89,9 per cent sensitivity and 97,4 per cent specificity, could be the test of choice for those laboratories which are equipped to perform ELISA. The "Leptoteste-S", with 89.9 per cent sensitivity and 94.8 per cent specificity, seems to be easier to perform and the most accessible to peripheral laboratories for rapid screening of human sera. Both techniques present the important characteristic of detecting early antibodies against leptospires, thus providing a diagnosis during the early stages of the disease.

Left ventricular maximal systolic elastance calculated by a combination of M-mode echocardiography and standard manometry

A method for obtaining the end-systolic left ventricular (LV) pressure-diameter and stress-diameter relationship in man was critically analyzed. Pressure-diameter and stress-diameter relationship were determined throughout the cardiac cycle by combining standard LV manometry with M-mode echocardiography. Nine adult patients with heart disease and without heart failure were studied during intracardiac catheterization under three different conditions of arterial pressure, i.e., basal (B) conditions (mean ñ SD systolic pressure, 102 ñ 10 mmHg) and two stable states of arterial hypertension (HI, 121 ñ 12 mmHg; HII, 147 ñ 17 mmHg) induced by venous infusion of phenylephrine after parasympathetic autonomic blockade with 0.04 mg/Kg atrophine. Significant reflex heart rate variation with arterial hypertension was observed (B, 115 ñ 20bpm; HI, 103 ñ 14 bpm; HII, 101 ñ 13 bpm) in spite of the parasympathetic blockade with atrophine. The linear end-systolic pressure-diameter and stress-diameter relationships ranged from 53.0 to 160.0 mmHg/cm and from 97.0 to 195.0 g/cm3, respectively. The end-systolic LV pressure-diameter and stress-diameter relationship lines presented high and variable slopes. The slopes, which are indicators of myocardial contractility, are susceptile to modifications by small deviations in the measurement of the ventricular diameter or by delay in the pressure curve recording

Ammiotic fluid lysozyme activity in fetal distress

Lysozyme activity was m,easured in amniotic fluid samples from 90 pregnant women with gestacional age ranging from 30 to 41 weeks. Twenty-nine samples were from high-risk subjects with different pathologies and signs of fetal distress. The control group consisted of 20 normal and 41 pathological pregnant women, whose disorders included Rh isoimmunization, diabetes, systemic arterial hypertension and pre-eclampsia without signs of fetal distress. amniotic fluid lysozyme levels in normal controls were similar to those detected in abnormal pregnant women without signs of fetal distress (x = 156.0 vs 131.8 microng/ml for 43-37 weeks of gestation), with a tendency toward higher values as pregnancy progressed to term in high-risk pregnant women with signs of fetal distress, regardless of neonate birth weight, than in subjects showing no such sugns (x = 40.3 and x = 25.4 microng/ml at 34-41 weeks of gestation, respectively). These data support the possibility of using amniotic fluid lysosyme activity levels as an indicator of fetal distress

Complemento hemolitico total e os componentes C3 e C7 em pacientes de paracoccidioidomicose. / Total hemolytic complement and C3 and C7 components in patients with paracoccidioidomycosis

Foram titulados o complemento hemolitico total e o componente C7 e dosado o componente C3 em 22 soros de pacientes com paracoccidioidomicose, 20 do sexo masculino e 2 do feminino, com idade entre 8 e 68 anos. Todos apresentavam a forma disseminada da molestia e tiveram a comprovacao do diagnostico clinico atraves de exames micologicos e imunologicos.O complemento hemolitico total variou entre 128 e 394 unidades (CH50) por ml de soro, com media (X) de 269,55 unidades/ml. O componente C7 variou entre 128 a 4096, em titulo expresso como a reciproca da diluicao do soro. O componente C3 variou entre 74 e 183 mg/100 ml de soro, com media (X) de 118,14 mg/100 ml.Os resultados obtidos nos pacientes de paracoccidioidomicose foram analisados estatisticamente, atraves do test "t" de Student para o complemento hemolitico total e componente C3 e pelo metodo do Qui-quadrado (em correcao de Yates) para o componente C7, comparando-os com os resultados obtidos em 30 individuos normais adultos, examinados concomitantemente. Tal analise permitiu concluir que nos pacientes de forma disseminada da paracoccidioidomicose o complemento hemolitico total e o componente C7 se encontravam dentro dos limites normais e o componente C3 se apresentava aumentado

Determinacao do complemento hemolitico total e do componente C3 em pacientes de esquistossomose mansonica. / Determination of total hemolytic complement and C3 component in patients with schistosomiasis mansoni

Foram estudadas 22 amostras de soro de pacientes com esquistossomose mansonica, com idades variando entre 6 a 46 anos, 8 do sexo masculino e 14do feminino, todos eles procedentes do ambulatorio de esquistossomose do Instituto de Medicina Tropical de Sao Paulo.O complemento hemolitico total variou de 115 a 284 unidades (CH50) por mililitro, media (X) de 209,23 unidades por ml e o componente C3 variou entre 50 e 136 mg/100ml, media de 93 50 mg/100ml. A analise estatistica indicou que o complemento hemolitico total se encontra diminuido enquanto que o componente C3 situa-se dentro dos limites normais, em casos de esquistossomose mansonica, para um erro de 1a. especie igual a 5%

Determinacao do complemento hemolitico total e do componente C3 em pacientes de meningite meningococica. / Determination of total hemolytic complement and of C3 component in patients with meningococcal meningitis

Foram estudadas 25 amostras de soro de pacientes com meningite meningococica, de idades compreendidas entre 1 e 57 anos, 16 do sexo masculino e 9 do feminino, internados no Hospital Emilio Ribas, durante surto epidemico, ocorrido em 1974, em Sao Paulo, Brasil. O diagnostico clinico teve sua comprovacao laboratorial atraves dos exames citologico, bioquimico e bacteriologico do liquor. Todos os soros foram colhidos nos primeiros cinco dias de molestia. Muitos pacientes apresentavam quadro clinico compativel com o diagnostico meningococcemia. O complemento hemolitico total variou de 69 a 383 unidades (CH50) por milimetro, media (X) de 213,88 unidades por ml e o componente C3 variou entre 11 e 116 mg/100 ml, media (X) de 78,52 mg/100 ml.A analise estatistica indicou diminuicao dos niveis sericos do complemento hemolitico total e do componente C3 em pacientes de meningite meningococica