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      HIV-associated tuberculous lymphadenitis: the importance of polymerase chain reaction (PCR) as a complementary tool for the diagnosis of tuberculosis - a study of 104 patients Translated title: Linfadenite tuberculosa associada ao HIV: a importância da reação em cadeia de polimerase (PCR) como ferramenta complementar para o diagnóstico da tuberculose - estudo de 104 pacientes

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          Abstract

          BACKGROUND: Lymphadenitis is common in HIV-positive patients. Diagnosis of the infections associated with this condition is complex, particularly in the case of tuberculosis. Rapid and specific detection of Mycobacterium tuberculosis (M. tuberculosis) is fundamental in ensuring adequate treatment. In addition, frequent causes of lymphadenitis such as those associated with lymphoma and histoplasmosis, among others, must be eliminated as possible causes. OBJECTIVES: To evaluate the accuracy of polymerase chain reaction as a tool for the diagnosis of lymphadenitis resulting from M. tuberculosis. METHODS: In this study, a protocol was developed using the following procedures: direct microscopy using Ziehl-Neelsen staining, culture in Lowenstein-Jensen medium, histology and polymerase chain reaction. RESULTS: A total of 104 patients were included in the study. According to histopathology, 38 patients (36%) were found to have nonspecific chronic lymphadenitis, 27 (26%) had tuberculous lymphadenitis, 11 patients (10.5%) had lymphoma and 9 (8.7%) had histoplasmosis. When Lowenstein-Jensen culture was performed, positive tests for tuberculous lymphadenitis increased by 30%. With polymerase chain reaction, M. tuberculosis DNA was detected in 6 out of 38 samples of non-specific chronic lymphadenitis. Three of these patients were followed up, developed symptoms of tuberculosis and were cured following specific treatment. CONCLUSION: The data obtained in this study suggest that all cases of lymphadenopathies should be submitted to histopathology, Lowenstein-Jensen or Ogawa culture and polymerase chain reaction. Polymerase chain reaction may prove to be useful in providing an early and accurate detection of cases of extrapulmonary tuberculosis in HIV-positive patients with lymphadenopathies, avoiding empirical treatment and the possible development of resistant strains.

          Translated abstract

          FUNDAMENTOS: A linfadenite é comum em pacientes HIV-positivos. O diagnóstico das infecções associadas a essa condição é complexo, especialmente em relação à tuberculose. A detecção rápida e específica do Mycobacterium tuberculosis (M. tuberculosis) é essencial para o tratamento adequado. Além disso, frequentes causas de linfadenites tais como as relacionadas a linfoma, histoplasmose, e outras, devem ser afastadas. OBJETIVOS: Avaliar a importância da biópsia cirúrgica excisional das linfadenites e realização dos exames histopatológicos, exames baciloscópicos (Ziehl-Neelsen), cultura (Lowenstein-Jensen) e reação em cadeia da polimerase (PCR) com finalidade diagnostica MÉTODOS: Desenvolveu-se um protocolo para biópsias excisionais de linfonodos, adotando-se os seguintes procedimentos: exame direto de esfregaços corados pelo método de Ziehl-Neelsen, cultura em meio de Lowenstein-Jensen, exame anátomo-patológico e PCR. RESULTADOS: Ao todo foram incluídos 104 pacientes, e o exame histopatológico evidenciou 36% (n = 38 pacientes) de casos de linfadenite crônica inespecífica, 26% (n = 27) de linfadenite tuberculosa, 10,5% (n =11) de linfoma e 8,7% (n = 9) de histoplasmose. Através da cultura no meio de Lowenstein-Jensen, os casos positivos para linfadenite tuberculosa aumentaram para 30%. A PCR detectou M. tuberculosis em 6 das 38 amostras de linfadenite crônica inespecífica. Três desses pacientes foram acompanhados, exibiram sintomas de tuberculose e foram curados após tratamento específico. CONCLUSÃO: Os dados obtidos neste trabalho sugerem que em todos os casos de linfadenopatia deve-se realizar exame histopatológico, cultura em Lowenstein-Jensen ou Ogawa e reação em cadeia da polimerase. A reação em cadeia da polimerase pode ser útil na detecção precoce e acurada de casos de TB extrapulmonar nos pacientes HIV-positivos com linfadenopatias, evitando-se o uso de tratamentos empíricos e o eventual desenvolvimento de cepas resistentes.

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          Extrapulmonary tuberculosis: an overview.

          In the 1980s, after a steady decline during preceding decades, there was a resurgence in the rate of tuberculosis in the United States that coincided with the acquired immunodeficiency syndrome epidemic. Disease patterns since have changed, with a higher incidence of disseminated and extrapulmonary disease now found. Extrapulmonary sites of infection commonly include lymph nodes, pleura, and osteoarticular areas, although any organ can be involved. The diagnosis of extrapulmonary tuberculosis can be elusive, necessitating a high index of suspicion. Physicians should obtain a thorough history focusing on risk behaviors for human immunodeficiency virus (HIV) infection and tuberculosis. Antituberculous therapy can minimize morbidity and mortality but may need to be initiated empirically. A negative smear for acid-fast bacillus, a lack of granulomas on histopathology, and failure to culture Mycobacterium tuberculosis do not exclude the diagnosis. Novel diagnostic modalities such as adenosine deaminase levels and polymerase chain reaction can be useful in certain forms of extrapulmonary tuberculosis. In general, the same regimens are used to treat pulmonary and extrapulmonary tuberculosis, and responses to antituberculous therapy are similar in patients with HIV infection and in those without. Treatment duration may need to be extended for central nervous system and skeletal tuberculosis, depending on drug resistance, and in patients who have a delayed or incomplete response. Adjunctive corticosteroids may be beneficial in patients with tuberculous meningitis, tuberculous pericarditis, or miliary tuberculosis with refractory hypoxemia.
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            Tuberculosis control and elimination 2010-50: cure, care, and social development.

            Rapid expansion of the standardised approach to tuberculosis diagnosis and treatment that is recommended by WHO allowed more than 36 million people to be cured between 1995 and 2008, averting up to 6 million deaths. Yet tuberculosis remains a severe global public health threat. There are more than 9 million new cases every year worldwide, and the incidence rate is falling at less than 1% per year. Although the overall target related to the Millennium Development Goals of halting and beginning to reverse the epidemic might have already been reached in 2004, the more important long-term elimination target set for 2050 will not be met with present strategies and instruments. Several key challenges persist. Many vulnerable people do not have access to affordable services of sufficient quality. Technologies for diagnosis, treatment, and prevention are old and inadequate. Multidrug-resistant tuberculosis is a serious threat in many settings. HIV/AIDS continues to fuel the tuberculosis epidemic, especially in Africa. Furthermore, other risk factors and underlying social determinants help to maintain tuberculosis in the community. Acceleration of the decline towards elimination of this disease will need invigorated actions in four broad areas: continued scale-up of early diagnosis and proper treatment for all forms of tuberculosis in line with the Stop TB Strategy; development and enforcement of bold health-system policies; establishment of links with the broader development agenda; and promotion and intensification of research towards innovations. Copyright 2010 Elsevier Ltd. All rights reserved.
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              Evaluation of real-time and conventional PCR targeting complex 85 genes for detection of Mycobacterium leprae DNA in skin biopsy samples from patients diagnosed with leprosy.

              In spite of the decrease in the number of registered leprosy patients, the number of new cases diagnosed each year (400,000) has remained essentially unchanged. Leprosy diagnosis is difficult due to the low sensitivity of current methodologies to identify new cases. In this study, conventional and TaqMan real-time PCR assays for detection of Mycobacterium leprae DNA were compared to current classification based on clinical, bacteriological, and histological evaluation. M. leprae DNA was extracted from frozen skin biopsy specimens from 69 leprosy patients enrolled in the study and was amplified using specific primers for either the antigen 85B-coding gene or the 85A-C intergenic region by using conventional and real-time PCR. The detection rate was 100% among multibacillary (MB) patients and ranged from 62.5% to 79.2% among paucibacillary (PB) patients according to the assay used. The TaqMan system for 85B gene amplification showed the highest sensitivity, although conventional PCR using the 85A-C gene as a target was also efficient. The cycle threshold (C(T)) values obtained using the TaqMan system were able to statistically (P < 0.0001) differentiate MB (mean C(T), 28.06; standard deviation [SD], 4.51) from PB (mean C(T), 33.06; SD, 2.24) patients. Also, there was a correlation between C(T) values and the bacteriological index for MB patients (Pearson's r, -0.444; P = 0.008). Within the PB patients' group, we tested normal skin from six patients exhibiting the pure neuritic form of leprosy (PNL). Five out of six PNL patients were positive for the presence of M. leprae DNA, even in the absence of skin lesions. In conclusion, the TaqMan real-time PCR developed here seems to be a useful tool for rapidly detecting and quantifying M. leprae DNA in clinical specimens in which bacilli were undetectable by conventional histological staining.
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                Author and article information

                Journal
                abd
                Anais Brasileiros de Dermatologia
                An. Bras. Dermatol.
                Sociedade Brasileira de Dermatologia (Rio de Janeiro, RJ, Brazil )
                0365-0596
                1806-4841
                October 2011
                : 86
                : 5
                : 925-931
                Affiliations
                [02] Manaus Amazonas orgnameAmazonas Foundation of Tropical Medicine Brazil
                [03] Rio de Janeiro orgnameOswaldo Cruz Foundation orgdiv1Oswaldo Cruz Institute orgdiv2Leprosy Laboratory Brazil
                [04] orgnameNilton Lins University
                [01] Manaus Amazonas orgnameNilton Lins University Center Brazil
                Article
                S0365-05962011000500010 S0365-0596(11)08600510
                61bff1bd-b297-4c25-b089-7e9b8d29420e

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 15 November 2010
                : 03 November 2010
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 29, Pages: 7
                Product

                SciELO Brazil

                Categories
                Investigation

                Tuberculose dos linfonodos,Tuberculose,Infecções oportunistas relacionadas com a AIDS,HIV,Tuberculosis, lymph node,Tuberculosis,AIDS-related opportunistic infections

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