DNA amplification by the polymerase chain reaction for the rapid diagnosis of tuberculous meningitis. Comparison of protocols involving three mycobacterial DNA sequences, IS6110, 65 kDa antigen, and MPB64

Lee, B.W.; Tan, Ji; Wong, Sek-Man; Tan, C. B.; Yap, Hui-Kim; Low, P. S.; Chia, Jer-Ming; Tay, J. S. H.

doi:10.1016/0022-510x(94)90220-8

Cited by 58 publications

(58 citation statements)

References 28 publications

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“…However, the true sensibility of PCR for tuberculosis in these samples may be higher, because the inclusion critérium (clinical and laboratory findings) was the probability one, not that of safety. There was no positive result in the second group of patients, suggesting a good specificity for this test 5 .…”

Section: Commentsmentioning

confidence: 81%

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Polymerase chain reaction in the diagnosis of tuberculous meningitis: preliminary report

Machado

Livramento

Bydlowski

et al. 1994

Arq. Neuro-Psiquiatr.

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In this preliminary report the results of PCR for detection of DNA sequences (65 KDa antigen) of Mycobacterium tuberculosis in CSF samples from 20 patients are registered. In 10 patients there were clinical and laboratory findings suggesting the diagnosis of tuberculous meningitis (test group). In the other 10 patients, clinical and laboratory findings suggested meningitis or meningo-encephalitis from other etiologies (control group). In 7 patients from the test group antigenic DNA sequences of Mycobacterium tuberculosis were found in CSF by PCR; positive results were not registered in the control group.

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Section: Commentsmentioning

confidence: 81%

“…The 383 bp sequence of the 65 KDa antigen of Mycobacterium tuberculosis was amplified 2,5 . After agarose gel electrophoresis, amplification products were transferred to nylon membrane by Southern blotting technique and hybridized with a specific M. tuberculosis probe 1 3 .…”

Section: Methodsmentioning

confidence: 99%

Polymerase chain reaction in the diagnosis of tuberculous meningitis: preliminary report

Machado

Livramento

Bydlowski

et al. 1994

Arq. Neuro-Psiquiatr.

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“…Many regions of the mycobacterial genome have been used in molecular diagnosis of TB, such as IS6110, 16S rRNA, hsp65, rpoB, sdaA, devR and mpt64 (Manjunath et al, 1991;Lachnik et al, 2002;Kim et al, 2004;Chakravorty et al, 2005;Negi et al, 2007;Hwang et al, 2013;Nimesh et al, 2013). mpt64 is found as a single copy in the genome of MTBC species (Lee et al, 1994), and has been used for both pulmonary and extrapulmonary TB diagnosis (Tan et al, 1995;Dar et al, 1998;Martins et al, 2000;Bhanu et al, 2005;Therese et al, 2005;Takahashi & Nakayama, 2006;Dil-Afroze et al, 2008;Kusum et al, 2012;Sethi et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of Mycobacterium tuberculosis complex by real-time PCR in sputum samples and its use in the routine diagnosis in a reference laboratory

Pinhata

Cergole-Novella²,

Carmo³

et al. 2015

Journal of Medical Microbiology

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Tuberculosis (TB) is an infectious disease of global distribution, constituting a serious public health problem in Brazil. Sã o Paulo State, located in the south-east of Brazil, notified 16 580 new TB cases in 2013. The Instituto Adolfo Lutz is a public health reference laboratory for TB diagnosis for all the State. Considering that rapid and accurate diagnosis is essential for TB control, the aim of this study was to evaluate the use of an in-house real-time (RT)-PCR assay targeting the mpt64 gene in the routine diagnosis of TB, and to compare this technique with smear microscopy and culture. From August 2012 to October 2013, 715 sputum samples from 657 patients were included in the study. Smear microscopy, culture, phenotypic and PRAhsp65 identification of mycobacteria, and mpt64 RT-PCR were performed. With respect to confirmed TB cases (n562/657; 9.4 %), smear microscopy had a sensitivity of 82.3 %. Culture and RT-PCR showed the same sensitivity, i.e. 90.3 %. Specificity was 99.7, 99.4 and 98.6 % for smear microscopy, culture and RT-PCR, respectively. mpt64 RT-PCR showed high sensitivity and specificity for the detection of Mycobacterium tuberculosis complex in sputum samples. This technique can be deployed in laboratories that do not have a rapid test for TB available, enabling the performance of TB diagnosis in up to 5 h.

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“…No result may be surely considered as falsepositive among those obtained with our nested PCR. Interestingly, a recent comparison 12 showed that a PCR targeting the MPB 64 gene was considerably more specific than reactions based on the 65 kDa heat shock protein (HSP) gene and the repeated sequence IS6110.…”

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confidence: 99%

Low sensitivity of polymerase chain reaction for diagnosis of tuberculous meningitis in southeastern Brazil

Brienze

et al. 2001

Rev. Soc. Bras. Med. Trop.

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Two polymerase chain reaction (PCR) protocols showed low sensitivity (36% and 53% for TB AMPLICOR and MPB64 nested PCR, respectively), when compared with classic microbiological methods (73% and 54% for Ziehl-Neelsen staining and culture, respectively), in the diagnosis of tuberculous meningitis in 91 patients in southeastern Brazil. Only three PCR-positive, microbiologically negative patients were found. Analysis of sequential cerebrospinal fluid samples by nested PCR detected Mycobacterium tuberculosis DNA up to 29 days after the introduction of antituberculosis chemotherapy. Key-words: Diagnosis of tuberculous meningitis. PCR. Mycobacterium tuberculosis.Resumo Dois protocolos de reação em cadeia da polimerase (PCR) apresentaram baixa sensibilidade (36% e 53%, respectivamente), para TB AMPLICOR e PCR aninhado baseado no gene MPB64), quando comparados aos métodos microbiológicos clássicos (73% e 54% respectivamente para baciloscopia e cultura), no diagnóstico de meningite tuberculosa em 91 pacientes do sudeste do Brasil. Somente três pacientes apresentaram PCR positiva e microbiologia negativa. A análise de amostras seqüenciais de líquor com a PCR aninhada detectou DNA de Mycobacterium tuberculosis até 29 dias após a introdução de tratamento. Palavras-chaves: Diagnóstico da meningite tuberculosa. PCR. Mycobacterium tuberculosis.

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DNA amplification by the polymerase chain reaction for the rapid diagnosis of tuberculous meningitis. Comparison of protocols involving three mycobacterial DNA sequences, IS6110, 65 kDa antigen, and MPB64

Cited by 58 publications

References 28 publications

Polymerase chain reaction in the diagnosis of tuberculous meningitis: preliminary report

Polymerase chain reaction in the diagnosis of tuberculous meningitis: preliminary report

Rapid detection of Mycobacterium tuberculosis complex by real-time PCR in sputum samples and its use in the routine diagnosis in a reference laboratory

Low sensitivity of polymerase chain reaction for diagnosis of tuberculous meningitis in southeastern Brazil

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