Department of internal medicine, Executive Yuan Department of health, Division of Respiratory and Critical Care Medicine, Taichung Hospital, Taichung, Taiwan.
BMC Microbiol. 2012 Jul 8;12:137. doi: 10.1186/1471-2180-12-137.
Mycobacteria can be quickly and simply identified by PCR restriction-enzyme analysis (PRA), but misidentification can occur because of similarities in band sizes that are critical for discriminating among species. Capillary electrophoresis can provide computer-aided band discrimination. The aim of this research was to develop an algorithm for identifying mycobacteria by combined rpoB duplex PRA (DPRA) and hsp65 PRA with capillary electrophoresis.
Three hundred and seventy-six acid-fast bacillus smear-positive BACTEC cultures, including 200 Mycobacterium tuberculosis complexes (MTC) and 176 non-tuberculous mycobacteria (NTM) were analyzed. With combined hsp65 and rpoB DPRA, the accuracy rate was 100% (200 isolates) for the MTC and 91.4% (161 isolates) for the NTM. Among the discordant results (8.6%) for the NTM, one isolate of Mycobacterial species and an isolate of M. flavescens were found as new sub-types in hsp65 PRA.
This effective and novel identification algorithm using combined rpoB DPRA and hsp65 PRA with capillary electrophoresis can rapidly identify mycobacteria and find new sub-types in hsp65 PRA. In addition, it is complementary to 16 S rDNA sequencing.
分枝杆菌可以通过 PCR 限制性内切酶分析（PRA）快速而简单地鉴定，但由于种间鉴定的关键带大小相似，可能会出现错误鉴定。毛细管电泳可以提供计算机辅助的带区分。本研究旨在开发一种通过组合 rpoB 双 PRA（DPRA）和 hsp65 PRA 与毛细管电泳鉴定分枝杆菌的算法。
分析了 376 例抗酸杆菌涂片阳性 BACTEC 培养物，包括 200 株结核分枝杆菌复合体（MTC）和 176 株非结核分枝杆菌（NTM）。采用组合 hsp65 和 rpoB DPRA，MTC 的准确率为 100%（200 株），NTM 为 91.4%（161 株）。在 NTM 的不一致结果（8.6%）中，发现一株分枝杆菌种和一株黄分枝杆菌在 hsp65 PRA 中为新亚型。
这种使用组合 rpoB DPRA 和 hsp65 PRA 与毛细管电泳的有效且新颖的鉴定算法可以快速鉴定分枝杆菌并在 hsp65 PRA 中发现新亚型。此外，它与 16S rDNA 测序互补。
