结合分枝杆菌生长指示管、罗氏COBAS AMPLICOR系统及双重聚合酶链反应，提高分枝杆菌的检测与鉴别能力。

Improved detection and differentiation of mycobacteria with combination of Mycobacterium Growth Indicator Tube and Roche COBAS AMPLICOR System in conjunction with Duplex PCR.

作者信息

Oh E J, Park Y J, Chang C L, Kim B K, Kim S M

机构信息

Department of Clinical Pathology, College of Medicine, the Catholic University of Korea, Kangnam St. Mary's Hospital, 505 Banpo-dong Seocho-ku, 137-040, Seoul, South Korea.

出版信息

J Microbiol Methods. 2001 Jul 30;46(1):29-36. doi: 10.1016/s0167-7012(01)00254-8.

DOI:10.1016/s0167-7012(01)00254-8

PMID:11412911

Abstract

In this study, a combination of liquid and solid media (current "gold standard" for culture) with combinations of liquid media (Mycobacteria Growth Indicator Tube (MGIT)) plus a commercial amplification system (Roche COBAS AMPLICOR System (CAS)), and solid media (Ogawa) plus CAS for detection of Mycobacterium tuberculosis were compared. In addition, the ability of the MGIT to recover mycobacteria from various clinical samples was compared with the abilities of egg-based Ogawa medium using equal volume of samples and a high concentration (6%) of NaOH for decontamination. A total of 705 specimens (395 respiratory and 310 extrapulmonary) that were collected from 554 patients were tested in parallel with three assays. The results of MGIT and Ogawa were evaluated with the "gold standard" (combination of culture and clinical data) and those of CAS were evaluated with extended gold standard including treated tuberculosis. A total of 130 mycobacterial infections (M. tuberculosis, n=122; mycobacterium other than tuberculosis (MOTT), n=8) were detected. The differentiation of M. tuberculosis and MOTT was successfully accomplished using duplex PCR. The overall sensitivity of the MGIT, Ogawa, and CAS for M. tuberculosis was 89.9%, 73.9%, and 79.9%, respectively. For the MOTT, the corresponding values for the MGIT and Ogawa medium were 100% and 12.5%, respectively. The mean detection time for M. tuberculosis was 22 days using MGIT and 32 days when using the Ogawa medium. The specificity of CAS was 98.4%, with an inhibition rate of 1.4%. A combination of MGIT plus CAS detected 97.5% of all M. tuberculosis infections (compared with MGIT plus Ogawa, 91.8%, P<0.05; compared with Ogawa plus CAS, 87.7%. P<0.01). Our results indicate that a combination of MGIT plus a Roche CAS in conjunction with duplex PCR, would be quite useful in clinical laboratories for both rapid detection and differentiation of M. tuberculosis and MOTT.

摘要

在本研究中，对液体和固体培养基的组合（当前培养的“金标准”）与液体培养基组合（分枝杆菌生长指示管（MGIT））加商业扩增系统（罗氏COBAS AMPLICOR系统（CAS））以及固体培养基（小川培养基）加CAS用于检测结核分枝杆菌的情况进行了比较。此外，使用等量样本和高浓度（6%）氢氧化钠进行去污处理，将MGIT从各种临床样本中回收分枝杆菌的能力与基于鸡蛋的小川培养基的能力进行了比较。从554名患者收集的总共705份标本（395份呼吸道标本和310份肺外标本）用三种检测方法进行了平行检测。MGIT和小川培养基的结果用“金标准”（培养和临床数据的组合）进行评估，CAS的结果用包括经治疗结核病的扩展金标准进行评估。总共检测到130例分枝杆菌感染（结核分枝杆菌，n = 122；非结核分枝杆菌（MOTT），n = 8）。使用双重PCR成功实现了结核分枝杆菌和MOTT的鉴别。MGIT、小川培养基和CAS对结核分枝杆菌的总体敏感性分别为89.9%、73.9%和79.9%。对于MOTT，MGIT和小川培养基的相应值分别为100%和12.5%。使用MGIT检测结核分枝杆菌的平均时间为22天，使用小川培养基时为32天。CAS的特异性为98.4%，抑制率为1.4%。MGIT加CAS的组合检测到所有结核分枝杆菌感染的97.5%（与MGIT加小川培养基相比为91.8%，P<0.05；与小川培养基加CAS相比为87.7%，P<0.01）。我们的结果表明，MGIT加罗氏CAS的组合结合双重PCR，在临床实验室中对于结核分枝杆菌和MOTT的快速检测和鉴别将非常有用。