Direct identification of mycobacteria from smear-positive sputum samples using an improved multiplex polymerase chain reaction assay

Ju Hsin Chia, Tsu Lan Wu, Lin Hui Su, An Jing Kuo, Hsin Chih Lai*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

8 Scopus citations

Abstract

The rapid identification of mycobacteria from smear-positive sputum samples is very important. To identify the . Mycobacterium tuberculosis complex (MTBC) and frequently isolated nontuberculous mycobacteria strains directly from smear-positive sputum samples, an improved multiplex polymerase chain reaction (PCR) assay was developed. Nine pairs of primers targeting the 16S-23S rDNA internal transcribed spacer-1, . hsp65, and the early secretory antigen (ESAT-6) gene sequences were developed, and their efficacy was evaluated in comparison to traditional culturing and 16S rRNA gene sequencing methods. A total of 200 smear- and culture-positive sputum specimens collected between November 2005 and May 2006 were used for the analysis. The results of the assay showed an accurate identification rate for acid-fast bacilli (AFB) 3+, AFB 2+, and AFB rare/1+ samples of 98%, 95%, and 53%, respectively. The improved multiplex PCR method saves time and has advantages for identifying mycobacteria from AFB 2+ and 3+ sputum samples. The method is suitable for use in countries with a high MTBC prevalence rate.

Original languageEnglish
Pages (from-to)340-349
Number of pages10
JournalDiagnostic Microbiology and Infectious Disease
Volume72
Issue number4
DOIs
StatePublished - 04 2012

Keywords

  • Direct identification
  • Multiplex PCR assay
  • Mycobacterium

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