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

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• 作者：Ju-Hsin Chia^a ; ^b ; ¹ ; Tsu-Lan Wu^a ; ^b ; ¹ ; Lin-Hui Su^a ; ^b ; An-Jing Kuo^a ; ^b ; Hsin-Chih Lai^b ; ^{hclai@mail.cgu.edu.tw}
• 关键词：Multiplex PCR assay ; Mycobacterium ; Direct identification
• 刊名：Diagnostic Microbiology and Infectious Disease
• 出版年：2012
• 期刊代码：79_07328893
• 类别：med
• 出版时间：April, 2012
• 卷：72
• 期：4
• 页码：340-349
• 文件大小：351 K

摘要

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.