Single-tube nested PCR assay with in-house DNA extraction for Mycobacterium tuberculosis detection in blood and urine
Rev. Soc. Bras. Med. Trop
; 48(6): 731-738, Nov.-Dec. 2015. tab
Article
en En
| LILACS
| ID: lil-767828
Biblioteca responsable:
BR1.1
ABSTRACT
Abstract INTRODUCTION:
Molecular analyses are auxiliary tools for detecting Koch's bacilli in clinical specimens from patients with suspected tuberculosis (TB). However, there are still no efficient diagnostic tests that combine high sensitivity and specificity and yield rapid results in the detection of TB. This study evaluated single-tube nested polymerase chain reaction (STNPCR) as a molecular diagnostic test with low risk of cross contamination for detecting Mycobacterium tuberculosis in clinical samples.METHODS:
Mycobacterium tuberculosis deoxyribonucleic acid (DNA) was detected in blood and urine samples by STNPCR followed by agarose gel electrophoresis. In this system, reaction tubes were not opened between the two stages of PCR (simple and nested).RESULTS:
STNPCR demonstrated good accuracy in clinical samples with no cross contamination between microtubes. Sensitivity in blood and urine, analyzed in parallel, was 35%-62% for pulmonary and 41%-72% for extrapulmonary TB. The specificity of STNPCR was 100% in most analyses, depending on the type of clinical sample (blood or urine) and clinical form of disease (pulmonary or extrapulmonary).CONCLUSIONS:
STNPCR was effective in detecting TB, especially the extrapulmonary form for which sensitivity was higher, and had the advantage of less invasive sample collection from patients for whom a spontaneous sputum sample was unavailable. With low risk of cross contamination, the STNPCR can be used as an adjunct to conventional methods for diagnosing TB.Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
LILACS
Asunto principal:
Tuberculosis
/
ADN Bacteriano
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Reacción en Cadena de la Polimerasa
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Mycobacterium tuberculosis
Tipo de estudio:
Diagnostic_studies
Límite:
Adolescent
/
Adult
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Aged
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Female
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Humans
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Male
Idioma:
En
Revista:
Rev. Soc. Bras. Med. Trop
Asunto de la revista:
MEDICINA TROPICAL
Año:
2015
Tipo del documento:
Article
/
Project document
País de afiliación:
Brasil