ABSTRACT Tuberculosis is an infectious disease of global importance with major economic and social burden accounting for 25% of all avoidable deaths in developing countries. Extrapulmonary involvement may occur either in association with clinically apparent pulmonary tuberculosis or in isolation. This cross-sectional descriptive study aimed to evaluate the impact of ocular tuberculosis in visual acuity at baseline and after two months of intensive anti-tuberculous therapy. A sample of 133 pulmonary tuberculosis patients, seven disseminated tuberculosis, and three pleural tuberculosis patients was evaluated. All patients underwent routine ophthalmic evaluation, including assessment of visual acuity, biomicroscopy, applanation tonometry, indirect ophthalmoscopy, and fluorescent angiography as appropriate. None of the patients had impaired visual acuity due to tuberculosis. A rate of 4.2% (6/143) of ocular involvement was found. None of the patients with ocular involvement were HIV-infected. Of the six patients with ocular involvement, five met the diagnostic criteria for probable and one for possible ocular lesions. As for the type of ocular lesions, two patients had bilateral findings: one had sclerouveitis and the second had choroidal nodules. The other four patients presented with unilateral lesions: peripheral retinal artery occlusion in the right eye (one case), choroidal nodules in the left eye (one case), and choroidal nodules in the right eye (two cases). Patients progressed favorably after two month of intensive therapy, with no significant reduction in vision.

OBJETIVO: Comparar quatro métodos laboratoriais no diagnóstico de tuberculose pulmonar. MÉTODOS: Foram realizadas pesquisa direta pelas colorações de Ziehl-Neelsen e auramina, cultura para micobactérias em meio Löwenstein-Jensen (LJ) e polymerase chain reaction (PCR, reação em cadeia da polimerase) para Mycobacterium tuberculosis em 160 amostras de secreção respiratória de pacientes com suspeita de tuberculose pulmonar. As cepas isoladas foram identificadas por método radiométrico utilizando-se p-nitro-alfa-acetilamino-beta-hidroxipropiofenona (NAP) e métodos clássicos. A sensibilidade dos métodos foi comparada com o padrão ouro para o diagnóstico da tuberculose pulmonar, definido por critérios clínicos, radiológicos e microbiológicos. RESULTADOS: Dos 160 pacientes, 142 foram diagnosticados com tuberculose pulmonar de acordo com o padrão ouro. As técnicas de Ziehl-Neelsen e auramina, cultura em meio LJ e PCR apresentaram sensibilidade de 54,2%, 58,4%, 67,6% e 77,5%, respectivamente, quando comparados ao critério diagnóstico adotado. A especificidade dos quatro métodos foi de 100%. A concordância na identificação da micobactéria entre PCR e o método radiométrico utilizando NAP foi alta (96,8%). A sensibilidade da PCR foi de 50,8% nas amostras com baciloscopia negativa e de 98,8% naquelas com baciloscopia positiva. Nas amostras com resultados negativos na baciloscopia e cultura, a sensibilidade da PCR foi menor que nas com resultados positivos (25,6% e 99,0%, respectivamente). CONCLUSÕES: A PCR é método promissor no diagnóstico da tuberculose pulmonar, mesmo em amostras paucibacilares. Além disso, apresenta a vantagem da identificação simultânea e rapidez do resultado.

OBJECTIVE: To compare four laboratory methods in the diagnosis of pulmonary tuberculosis. METHODS: Respiratory secretion specimens were collected from 160 patients suspected of having pulmonary tuberculosis. Direct testing for Mycobacterium tuberculosis was carried out using Ziehl-Neelsen and auramine staining. In addition, culture in Löwenstein-Jensen (LJ) medium and polymerase chain reaction (PCR) were used. The strains isolated were identified by means of a radiometric method using p-nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) and classical methods. The sensitivity of the methods was compared to the gold standard for the diagnosis of pulmonary tuberculosis, based on clinical, radiological and microbiological criteria. RESULTS: Of the 160 patients, 142 were diagnosed with pulmonary tuberculosis according to the gold standard. The sensitivity of Ziehl-Neelsen staining, auramine staining, culture in LJ medium and PCR was 54.2%, 58.4%, 67.6% and 77.5%, respectively, when compared with the diagnostic criterion adopted. All four methods presented 100% specificity. In the identification of mycobacteria, there was high (96.8%) concordance between PCR and the radiometric method using NAP. The sensitivity of PCR was 50.8% in samples with negative sputum smear microscopy results and 98.8% in those with positive results. The sensitivity of PCR was lower in specimens with negative results in sputum smear microscopy and culture than in those with positive results (25.6% and 99.0%, respectively). CONCLUSIONS: We found PCR to be a promising method for the diagnosis of pulmonary tuberculosis, even in paucibacillary specimens. Simultaneous identification and faster results are additional advantages of this method.

We evaluated the ability of a PCR assay to identify Mycobacterium tuberculosis complex (MTBC) from positive BACTEC® 12B broth cultures. A total of 107 sputum samples were processed and inoculated into Ogawa slants and BACTEC® 12B vials. At a growth index (GI) > 30, 1.0 ml of the 12B broth was removed, stored, and assayed with PCR. Molecular results were compared to those obtained by phenotypic identification methods, including the BACTEC® NAP method. The average times required to perform PCR and NAP were compared. Of the 107 broth cultures evaluated, 90 were NAP positive, while 91 were PCR positive for MTBC. Of particular interest were three contaminated BACTEC® 12B broth cultures yielding microorganisms other than acid-fast bacilli growth with a MTBC that were successfully identified by PCR, resulting in a mean time of 14 days to identify MTBC before NAP identification. These results suggest that PCR could be used as an alternative to the NAP test for the rapid identification of MTBC in BACTEC® 12B cultures, particularly in those that contained both MTBC and nontuberculous mycobacteria.

OBJETIVO: Desenvolver um sistema para o diagnóstico molecular da tuberculose por reação em cadeia da polimerase, do inglês polymerase chain reaction (PCR), pela construção de primers baseados na diferença da organização de uma região intergênica da fosfolipase (phospholipase) C (região plcB-plcC), que diferencia Mycobacterium tuberculosis das outras micobactérias. MÉTODOS: Um produto de PCR com o tamanho esperado (432 pb) foi obtido somente de M. tuberculosis e M. africanum. Um total de 33 isolados micobacterianos e 273 amostras clínicas de pacientes com suspeita de tuberculose foram examinados. Estes foram submetidos ao estudo comparativo da técnica de PCR contra o cultivo. RESULTADOS: Os dados mostraram 93,8% de exatidão para PCR contra o cultivo (p < 0,0001), 93,1% de sensibilidade (IC: 88,7-96,0) e especificidade de 96,4% (IC: 96,1-99,4). O valor de Kappa foi de 0,82, demonstrando um alinhamento perfeito para a verificação do grau de concordância entre os testes. CONCLUSÃO: O uso da região plcB-plcC para a amplificação por PCR é mostrado como uma ferramenta importante e de confiança para o diagnóstico específico de tuberculose nas amostras clínicas analisadas.

OBJECTIVE: To develop a system for the molecular diagnosis of tuberculosis by polymerase chain reaction (PCR), constructing primers based on the difference in gene organization of the intergenic region of phospholipase C (plcB-plcC region), which differentiates Mycobacterium tuberculosis from other mycobacteria. METHODS: A PCR product of the expected size (432 bp) was obtained from M. tuberculosis and M. africanum only. A total of 33 mycobacterial isolates and 273 clinical samples from patients suspected of having tuberculosis were examined. These were used in the comparative study of the PCR technique versus culture. RESULTS: For PCR versus culture, the data showed 93.8% accuracy (p < 0.0001), 93.1% sensitivity (CI: 88.7-96.0), and 96.4% specificity (CI: 96.1-99.4). The Kappa value (0.82) shows that there was a near-perfect concordance between the two tests. CONCLUSION: The use of the plcB-plcC region in PCR amplification was found to be an important and reliable tool for the specific diagnosis of tuberculosis in the samples analyzed.

Estudo retrospectivo de perfil dos casos de TBMR do Espírito Santo, entre 2000 e 2004. Identificou-se 61 pacientes com TBMR, sendo a amostra composta por 57 casos, que tiveram seus prontuários revisados para obtenção das variáveis estudadas. Estimou-se como prevalência para a TBMR combinada 0,87% (0,66 - 1,13; IC95%). O teste anti-HIV foi feito em 78,9% dos casos, sendo 11,1% positivos. Investigando co-morbidades, destacaram-se etilismo e tabagismo. Encontrou-se 11 casos de resistência primária (RI) e 46 de resistência adquirida (RA), com média de 2,3 ± 1,3 tratamentos anteriores. Em 35,1% dos casos houve relato de contato prévio conhecido com doente de tuberculose, enquanto em 67,9% não houve ou nega-se ter havido. Dez pacientes (17,5%) foram tratados com o esquema de 1ª linha, 18 (31,6%) com o de 2ª linha e 27 (47,4%) com o de 3ª linha. Dezoito (31,6%) tiveram tratamento auto-administrado, e 39 (68,4%) supervisionado. Quanto ao desfecho, houve cura em 33 casos (71,7%), abandono em 7 (15,2%) e óbito em 5 (10,9%), 1 caso de falência e 11 (19,3%) estão em tratamento. Dos 10 casos encerrados de RI, 80% (8/10) foram curados, contra 69,4% (25/36) dos casos de RA. Concluímos que a prevalência de TBMR é baixa no ES. A cura foi alcançada em 70% dos casos. As co-morbidades podem ser importante fator interveniente para um desfecho satisfatório do tratamento. Os resultados enfatizam a necessidade de busca, diagnóstico e realização de TSA para identificação da RI e a supervisão do tratamento de todos os casos de tuberculose.

This is a retrospective profile study of MRTB cases in Espírito Santo, between 2000 and 2004. Sixty-one patients were identified as MRTB, in a sample of 57 cases. All clinical charts of these cases were reviewed for the variables studied. The combined prevalence of MRTB was estimated as 0.87% (0.66 - 1.13; 95% CI). Anti-HIV tests were performed in 78.9% of cases, yielding 5 (11.1%) positive results. Regarding co-morbidity, alcoholism and smoking have a special importance. 11 cases (19.3%) of primary resistance (IR) were found, while 46 (80.7%) were cases of acquired resistance (AR), with an average of 2.3 ± 1.3 previous treatments. 35.1% of the cases informed a known previous contact with a person with tuberculosis, while for 67.9%, the contact was denied or was not the case. 10 patients (17.5%) had been treated with a 1st line regimen, 18 (31.6%) with a 2nd line one, and 27 (47.4%) with 3rd line. Eighteen (31.6%) had had self-administered treatment, while 39 (68.4%) had undergone supervised treatment. Outcome for 33 cases (71.7%) was cure, abandon for 7 (15.2%) and death for 5 (10.9%); there was one case of treatment failure and 11 (19.3%) were still in treatment. Of the 10 cases of IR with a defined outcome, 80% (8/10) were cured, against 69.4% (25/36) of AR cases. We concluded that the prevalence of MRTB is low in the state of Espírito Santo. Cure was reached in about 70% of all treatments. Associated co-morbidity can be a major drawback for satisfactory treatment outcome. Our results emphasize the need for case finding, search, diagnosis and performance of SAT for the identification of IR and implementation of supervised treatment for all tuberculosis cases.

Estudo retrospectivo de perfil dos casos de TBMR do Espírito Santo, entre 2000 e 2004. Identificou-se 61 pacientes com TBMR, sendo a amostra composta por 57 casos, que tiveram seus prontuários revisados para obtenção das variáveis estudadas. Estimou-se como prevalência para a TBMR combinada 0,87% (0,66 - 1,13; IC95%). O teste anti-HIV foi feito em 78,9% dos casos, sendo 11,1% positivos. Investigando co-morbidades, destacaram-se etilismo e tabagismo. Encontrou-se 11 casos de resistência primária (RI) e 46 de resistência adquirida (RA), com média de 2,3 ± 1,3 tratamentos anteriores. Em 35,1% dos casos houve relato de contato prévio conhecido com doente de tuberculose, enquanto em 67,9% não houve ou nega-se ter havido. Dez pacientes (17,5%) foram tratados com o esquema de 1ª linha, 18 (31,6%) com o de 2ª linha e 27 (47,4%) com o de 3ª linha. Dezoito (31,6%) tiveram tratamento auto-administrado, e 39 (68,4%) supervisionado. Quanto ao desfecho, houve cura em 33 casos (71,7%), abandono em 7 (15,2%) e óbito em 5 (10,9%), 1 caso de falência e 11 (19,3%) estão em tratamento. Dos 10 casos encerrados de RI, 80% (8/10) foram curados, contra 69,4% (25/36) dos casos de RA. Concluímos que a prevalência de TBMR é baixa no ES. A cura foi alcançada em 70% dos casos. As co-morbidades podem ser importante fator interveniente para um desfecho satisfatório do tratamento. Os resultados enfatizam a necessidade de busca, diagnóstico e realização de TSA para identificação da RI e a supervisão do tratamento de todos os casos de tuberculose.

This is a retrospective profile study of MRTB cases in Espírito Santo, between 2000 and 2004. Sixty-one patients were identified as MRTB, in a sample of 57 cases. All clinical charts of these cases were reviewed for the variables studied. The combined prevalence of MRTB was estimated as 0.87% (0.66 - 1.13; 95% CI). Anti-HIV tests were performed in 78.9% of cases, yielding 5 (11.1%) positive results. Regarding co-morbidity, alcoholism and smoking have a special importance. 11 cases (19.3%) of primary resistance (IR) were found, while 46 (80.7%) were cases of acquired resistance (AR), with an average of 2.3 ± 1.3 previous treatments. 35.1% of the cases informed a known previous contact with a person with tuberculosis, while for 67.9%, the contact was denied or was not the case. 10 patients (17.5%) had been treated with a 1st line regimen, 18 (31.6%) with a 2nd line one, and 27 (47.4%) with 3rd line. Eighteen (31.6%) had had self-administered treatment, while 39 (68.4%) had undergone supervised treatment. Outcome for 33 cases (71.7%) was cure, abandon for 7 (15.2%) and death for 5 (10.9%); there was one case of treatment failure and 11 (19.3%) were still in treatment. Of the 10 cases of IR with a defined outcome, 80% (8/10) were cured, against 69.4% (25/36) of AR cases. We concluded that the prevalence of MRTB is low in the state of Espírito Santo. Cure was reached in about 70% of all treatments. Associated co-morbidity can be a major drawback for satisfactory treatment outcome. Our results emphasize the need for case finding, search, diagnosis and performance of SAT for the identification of IR and implementation of supervised treatment for all tuberculosis cases.

O método de amplificação de DNA baseado na reação em cadeia da ligase (Abbott LCx MTB) foi avaliado para detecção do Mycobacterium tuberculosis em espécimes pulmonares. Os resultados do LCx MTB foram comparados aos resultados de baciloscopia, cultura e diagnóstico clínico para cada paciente. Um total de 297 espécimes (escarro e lavado broncoalveolar) de 189 pacientes foram testadas. Os valores de sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo do LCX vs cultura foram 92,7%, 93%, 67,8% e 98,7%, respectivamente. Quando comparados ao diagnóstico clínico, os valores de sensibilidade, especificidade, VPP e VPN para o LCx foram 88,9%, 96,8%, 86,5% e 97,4%, respectivamente. A sensibilidade do LCx MTB foi de 75% para as amostras com baciloscopia negativa e cultura positiva. Os resultados indicam que o teste LCx MTB é simples, rápido, eficiente e pode ser utilizado como um recurso complementar para o diagnóstico da tuberculose.

A ligase chain reaction DNA amplification method for direct detection of Mycobacterium tuberculosis (Abbott LCx MTB) in respiratory specimens was evaluated. Results from LCx MTB Assay were compared with those from acid fast bacilli smear, culture, and final clinical diagnosis for each patient. A total of 297 respiratory specimens (sputum and bronchial lavage) from 193 patients were tested. The sensitivity, specificity, positive predictive value and negative predictive value of LCx vs culture were 92.7%, 93%, 67.8% and 98.7%, respectively. When compared to the clinical final diagnosis, the sensitivity, specificity, PPV and NPV for LCx were 88.9%, 96.8%, 86.5% and 97.4%, respectively. The sensitivity of LCx MTB assay was 75% for smear-negative, culture positive samples. The results indicate that LCx MTB assay is a rapid, simple and valuable technique as a complementary tool for the diagnosis of tuberculosis.

The purpose of this study was to assess the distribution of Mycobacterium avium serovars isolated from AIDS patients in São Paulo and Rio de Janeiro. Ninety single site or multiple site isolates from 75 patients were examined. The most frequent serovars found were 8 (39.2%), 4 (21.4%) and 1 (10.7%). The frequency of mixed infections with serovar 8 or 4 was 37.8%. Among the 90 strains examined, M. intracellulare serovars (7 strains) and M. scrofulaceum (4 strains) were found in 11 isolates (12%) indicating that M. avium (88%) was the major opportunistic species in the M. avium complex isolates in Brazilian AIDS patients

Anterior a pandemia de AIDS, o Complexo Mycobacterium avium (MAC) era responsável pela maioria das vezes, por pneumopatias acometendo pacientes com doença pulmonar crônica de base como enfisema e bronquite crônica36. Em 1981, com o advento da síndrome de imunodeficiência adquirida (SIDA), o MAC passou a representar uma das doenças bacterianas mais frequentes em pacientes com esta síndrome, sendo a doença disseminada a principal forma de manifestação clínica da infecção8. Entre Janeiro de 1989 e Fevereiro de 1991, no Setor de Micobactérias do Instituto Adolfo Lutz em São Paulo, o MAC foi isolado de 103 pacientes a partir do cultivo de diferentes espécimes estéreis e não estéreis processados, coletados de 2.304 pacientes atendidos no Centro de Referência e Treinamento AIDS e/ou Instituto de Infectologia Emilio Ribas. A doença disseminada foi diagnosticada em 29 destes, com base no isolamento do MAC a partir do sangue e/ou aspirado de medula óssea. Os outros 74 pacientes foram agrupados nas categorias altamente (5), moderadamente (26) e pouco sugestiva de doença (43) de acordo com os postulados de DAVIDSON (1989)10. Os diferentes critérios para valorizar o seu isolamento de espécimes estéreis e não estéreis são discutidos.

Before the AIDS pandemia, the Mycobacterium avium complex (MAC) was responsible in most cases for the pneumopathies that attack patients with basic chronic pulmonary diseases such as emphysema and chronic bronchitis36. In 1981, with the advent of the acquired immunodeficiency syndrome (AIDS), MAC started to represent one of the most frequent bacterial diseases among AIDS patients, with the disseminated form of the disease being the major clinical manifestation of the infection8. Between January 1989 and February 1991, the Section of Mycobacteria of the Adolfo Lutz Institute, São Paulo, isolated MAC from 103 patients by culturing different sterile and no-sterile processed specimens collected from 2304 patients seen at the AIDS Reference and Training Center and/or Emilio Ribas Infectology Institute. Disseminated disease was diagnosed in 29 of those patients on the basis of MAC isolation from blood and/or bone marrow aspirate. The other 74 patients were divided into categories highly (5), moderately (26) and little suggestive of disease (43) according to the criteria of DAVIDSON (1989)10. The various criteria for MAC isolation from sterile and non-sterile specimens are discussed.

Amostras de líquido cefalorraquidiano (LCR) de 2083 pacientes com Síndrome de Imunodeficiência Adquirida (SIDA) e complicações neurológicas foram examinados durante um período de 7 anos (1984-1990). A porcentagem de pacientes que tiveram pelo menos um agente bacteriano cultivado do LCR foi de 6,2%. Mycobacterium tuberculosis foi o mais frequentemente isolado (4,3%), seguido do complexo Mycobacterium avium ou MAC (0,7%), de Pseudomonas spp (0,5%), Enterobacter spp (0,4%), e Staphylococcus aureus (0,3%). Entre 130 pacientes com cultura positiva, de 89 (68,5%) foi isolado M. tuberculosis e de 15 (11,6%) MAC. A frequência de isolamentos bacterianos aumentou de 1988 (5,2%) a 1990 (7,2%), particularmente devido ao maior isolamento de MAC. Os agentes bacterianos foram mais frequentemente isolados de pacientes na faixa etária de 21-30 anos e de mulheres (p<0.05).

Cerebrospinal fluid (CSF) samples from 2083 patients with acquired immunodeficiency syndrome (AIDS) and neurological complications were bacteriologically examined during a period of 7 years (1984-1990). The percentage of patients who had at least one bacterial agent cultured from the CSF was 6.2%. Mycobacterium tuberculosis was the most frequently isolated agent (4.3%), followed by Mycobacterium avium complex or MAC (0.7%), Pseudomonas spp (0.5%), Enterobacter spp (0.4%), and Staphylococcus aureus (0.3%). Among 130 culture positive patients, 89 (68.5%) had M. tuberculosis and 15 (11.6%) had MAC. The frequency of bacterial isolations increased from 1988 (5.2%) to 1990 (7.2%), partly due to the increase in MAC isolations. Bacterial agents were more frequently isolated from patients in the age group 21-30 years and from women (p<0.05).