A multiplex polymerase chain reaction assay for genus-, group- and species-specific detection of mycobacteria

被引：15

作者：

Kurabachew, M ^{[1
]}

Enger, O

Sandaa, RA

Skuce, R

Bjorvatn, B

机构：

[1] Univ Bergen, Ctr Int Hlth, N-5021 Bergen, Norway

[2] Univ Bergen, Dept Med Microbiol & Immunol, Gade Inst, N-5021 Bergen, Norway

[3] Univ Addis Ababa, Dept Biol, Addis Ababa, Ethiopia

[4] Forninova AS, N-5008 Bergen, Norway

[5] UiB, Dept Microbiol, N-5020 Bergen, Norway

[6] Dept Agr & Rural Dev NI, Vet Sci Div, Dept Bacteriol, Belfast BT4 3SD, Antrim, North Ireland

来源：

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE | 2004年 / 49卷 / 02期

关键词：

multiplex PCR; mycobacteria; oxyR gene; rDNA;

D O I：

10.1016/j.diagmicrobio.2004.03.010

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

We developed and evaluated a single-step, multiplex polymerase chain reaction (PCR) assay for distinguishing (1) between the Mycobacterium tuberculosis complex (MTBC) and mycobacteria other than tuberculosis (MOTT) and (2) between M. tuberculosis and M. bovis species. The assay targeted the 16S and the 23S rDNA to distinguish between MTBC and MOTT species, and the oxyl? gene to distinguish between M. tuberculosis and M. bovis strains. Clinical samples and reference strains (N = 156) comprised 93 strains of M. tuberculosis, 44 of M. bovis, 1 M. africanum strain, and 18 strains representing 9 different species of MOTTs. MOTTs generated only a single PCR product of about 2.5 kilobase; however, all of the MTBC strains produced a 118 base pair (bp) fragment and an additional 270 bp fragment was obtained for M. tuberculosis and M. qfricanum when the primer pair oxyRTl3-2.l1oxyRMT-l was used. When oxyRTB2.1 loxyRMB-1 primers were used, the 270 bp fragment was obtained for only M. bovis. The assay needed as little as 1 pg of purified genomic DNA to make a positive identification. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：99 / 104

页数：6

共 22 条

[1] Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species [J].