Cas12a/guide RNA-based platform for rapid and accurate identification of major Mycobacterium species
Cas12a/guide RNA-based platform for rapid and accurate identification of major Mycobacterium species
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基于 Cas12a/guide RNA 的平台，可快速准确地鉴定主要分枝杆菌物种
DOI：
10.1128/jcm.01368-19
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Guoliang Zhang
中科院分区：
文献类型：
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作者：
Guohui Xiao;Xing He;Su Zhang;Yaya Liu;Zhihang Liang;Houming Liu;Juanjuan Zhang;Min Ou;Shuhao Cai;Wenjie Lai;Tianyu Zhang;Lili Ren;Guoliang Zhang
结核分枝杆菌感染和非结核分枝杆菌（NTM）感染表现出相似的临床症状；然而，这两类感染的治疗方法是不同的。因此，快速、准确地鉴定结核分枝杆菌和NTM菌种对于控制结核病和NTM感染非常重要。在本研究中，开发了基于 Cas12a/引导 RNA (gRNA) 的平台来识别结核分枝杆菌和大多数 NTM 物种。通过设计针对 therpoB 序列的物种特异性 gRNA 探针，基于 Cas12a/gRNA 的平台成功鉴定了结核分枝杆菌和六种主要 NTM 物种（脓肿分枝杆菌、胞内分枝杆菌、鸟分枝杆菌、堪萨斯分枝杆菌、戈登分枝杆菌和偶然分枝杆菌），且没有交叉反应。在盲法评估中，73株临床分枝杆菌分离株中共有72株被正确鉴定，这与之前的rpoB测序结果一致。这些结果表明，基于 Cas12a/gRNA 的平台是快速、准确且经济有效地鉴定结核分枝杆菌和 NTM 物种的有前途的工具。
Mycobacterium tuberculosis infection and nontuberculous mycobacteria (NTM) infections exhibit similar clinical symptoms; however, the therapies for these two types of infections are different. Therefore, the rapid and accurate identification of M. tuberculosis and NTM species is very important for the control of tuberculosis and NTM infections. In the present study, a Cas12a/guide RNA (gRNA)-based platform was developed to identify M. tuberculosis and most NTM species. By designing species-specific gRNA probes targeting therpoBsequence, a Cas12a/gRNA-based platform successfully identified M. tuberculosis and six major NTM species (Mycobacterium abscessus, Mycobacterium intracellulare, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium gordonae, and Mycobacterium fortuitum) without cross-reactivity. In a blind assessment, a total of 72 out of 73 clinicalMycobacteriumisolates were correctly identified, which is consistent with previousrpoBsequencing results. These results suggest that the Cas12a/gRNA-based platform is a promising tool for the rapid, accurate, and cost-effective identification of both M. tuberculosis and NTM species.