A utilidade da biologia molecular no diagnóstico da tuberculose

Mello, Fernanda Carvalho de Queiroz; Fonseca-Costa, Joseane

doi:10.1590/s1806-37132005000300002

Cited by 10 publications

(6 citation statements)

References 5 publications

(5 reference statements)

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“…Over the years several studies proposed and validated molecular techniques for diagnosis of TBM [ 8 – 14 ]. These studies considered the importance of developing a simple technique that was easily reproduced in laboratories with minimal resources [ 10 , 11 , 13 , 15 , 16 ]. The choice of the most appropriate extraction method and the target to be amplified are criteria that improve the precision, sensitivity and specificity of the PCR test [ 16 – 18 ].…”

Section: Discussionmentioning

confidence: 99%

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Comparison of DNA Extraction Protocols and Molecular Targets to Diagnose Tuberculous Meningitis

Palomo

Rivero

Quiles

et al. 2017

Tuberculosis Research and Treatment

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Tuberculous meningitis (TBM) is a severe form of extrapulmonary tuberculosis. The aims of this study were to evaluate in-house molecular diagnostic protocols of DNA extraction directly from CSF samples and the targets amplified by qPCR as an accurate and fast diagnosis of TBM. One hundred CSF samples from 68 patients suspected of TBM were studied. Four DNA extraction techniques (phenol-chloroform-thiocyanate guanidine, silica thiocyanate guanidine, resin, and resin with ethanol) were compared and CSF samples were used to determine the best target (IS6110, MPB64, and hsp65 KDa) by qPCR. The extraction protocol using the phenol-chloroform-thiocyanate guanidine showed the best results in terms of quantification and sensitivity of PCR amplification, presenting up to 10 times more DNA than the second best protocol, the silica guanidine thiocyanate. The target that showed the best result for TBM diagnosis was the IS6110. This target showed 91% sensitivity and 97% specificity when we analyzed the results by sample and showed 100% sensitivity and 98% specificity when we analyzed the results by patient. The DNA extraction with phenol-chloroform-thiocyanate guanidine followed by IS6110 target amplification has been shown to be suitable for diagnosis of TBM in our clinical setting.

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Section: Discussionmentioning

confidence: 99%

“…The techniques using phenol-chloroform, silica, and thiocyanate guanidine are described as excellent choices for DNA extraction in different biological materials, in addition to being inexpensive and simple [ 8 – 13 ].…”

Section: Discussionmentioning

confidence: 99%

Comparison of DNA Extraction Protocols and Molecular Targets to Diagnose Tuberculous Meningitis

Palomo

Rivero

Quiles

et al. 2017

Tuberculosis Research and Treatment

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“…A reação em Cadeia Polimerase (PCR), foi desenvolvida nos anos 80 por Kary Mullis, um bioquímico estadunidense que ganhou o prêmio Nobel de química em 1993 pela sua descoberta, que revolucionou o mundo da pesquisa científica (MELLO, 2005).…”

Section: Reação Em Cadeia Da Polimerase (Pcr)unclassified

A importância da Biologia Molecular no diagnóstico do SARS-CoV-2

VELOZO,

PEIXOTO

2024

Rev. Cient. Multidisc. Saber

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The pandemic caused by the etiologic agent SARS-CoV- 2, started at the beginning of 2020. And create a big challenge in the scientific community from all around the world. The symptoms can variate of levels manifestation to severs cases of pneumonia. Studies are seeking ways of prevention, diagnosis and treatment of the disease. In view of the pandemic scenario and the worsening number of cases and deaths due to the disease, the search for an accurate diagnosis is essential. Therefore, the aim of this study is demonstrating the importance of using molecular biology through the technique of Real Time Transcriptase Reverse Polymerase Chain Reaction (Real Time RT-PCR), for the detection of the genetic material of SARS-CoV-2 from data found in the literature. After a critical analysis of the literature, it was possible to confirm the importance and efficiency of using the Real Time RT-PCR technique in the diagnosis of infection, since this technique allows a good performance in the detection of viral genetic material, guaranteeing a high success rate when differentiating an infected patient from the non-infected. Concluding that until now this technique is considered as the main diagnostic method for the disease, remaining as a “gold standard”.

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“…Desenvolvida por Kary Mullis, a técnica de reação em cadeia da polimerase ou PCR (do inglês, Polymerase Chain Reaction) pode amplificar uma região de DNA por ciclos repetidos de síntese de DNA, in vitro, em poucas horas (SAIKI et al, 1985;MELLO;FONSECA-COSTA, 2005). A PCR possui enorme aplicação na área forense e criminalística pelo fato de se tornar uma grande ferramenta na investigação de marcadores moleculares e permitir analisar casos de identidades contestáveis utilizando até mesmo sequências de DNA degradadas, ou fontes (OLIVEIRA et al, 2007;OLIVEIRA, J., 2019).…”

Section: Reação Em Cadeia Da Polimerase E Eletroforese Capilarunclassified