Liu Shuang, Huang Guangtao, Gong Yali, Jin Xiaojun, Meng Yudan, Peng Yizhi, Zhao Junning, Li Xiaolu, Li Qin
Department of Plastic & Burns Surgery, The Affiliated Hospital of Southwest Medical University, Tai Ping Street, Luzhou, 646000, China.
Institute of Burn Research, Southwest Hospital, The Army Medical University, Gao Tan Yan Street, Chongqing, 400038, China.
Burns Trauma. 2020 Sep 2;8:tkaa026. doi: 10.1093/burnst/tkaa026. eCollection 2020.
(A. ) is one of the pivotal pathogens responsible for nosocomial infections, especially in patients with low immune response, and infection with carbapenem-resistant has been increasing in recent years. Rapid and accurate detection of carbapenem-resistance genes in could be of immense help to clinical staff.
In this study, a 15-μL reaction system for recombinase polymerase amplification (RPA) was developed and tested. We collected 30 clinical isolates of from the Burn Institute of Southwest Hospital of Third Military Medical University (Army Medical University) for 6 months and tested antibiotic susceptibility using the VITEK 2 system. was detected based on the gene by PCR, qPCR and 15 μL-RPA, respectively. Sensitivity and specificity were evaluated. In addition, PCR and 15 μL-RPA data for detecting the carbapenem-resistance gene were comparatively assessed.
The detection limit of the gene by 15 μL RPA was 2.86 CFU/ml, with sensitivity comparable to PCR and qPCR. No positive amplification signals were detected in non- isolates, indicating high specificity. However, only 18 minutes were needed for the 15 μL RPA assay. Furthermore, an antibiotic susceptibility test showed that up to 90% of strains were resistant to meropenem and imipenem; 15 μL RPA data for detecting showed that only 10% ( = 3) of isolates did not show positive amplification signals, and the other 90% of ( = 27) isolates were positive, corroborating PCR results.
We demonstrated that the new 15 μL RPA assay for detecting in is faster and simpler than qPCR and PCR. It is a promising alternative molecular diagnostic tool for rapid and effective detection of and drug-resistance genes in the field and point-of-care testing.
（某菌）是医院感染的关键病原体之一，尤其是在免疫反应低下的患者中，近年来耐碳青霉烯类（该菌）的感染一直在增加。快速准确地检测该菌中的耐碳青霉烯类基因对临床工作人员有极大帮助。
在本研究中，开发并测试了一种用于重组酶聚合酶扩增（RPA）的15微升反应体系。我们从陆军军医大学第三军医大学西南医院烧伤研究所收集了6个月内的30株该菌临床分离株，并用VITEK 2系统检测抗生素敏感性。分别通过PCR、qPCR和15微升RPA基于（某基因）检测该菌。评估敏感性和特异性。此外，对检测耐碳青霉烯类基因（某基因）的PCR和15微升RPA数据进行比较评估。
15微升RPA对（某基因）的检测限为2.86 CFU/ml，敏感性与PCR和qPCR相当。在非（该菌）分离株中未检测到阳性扩增信号，表明特异性高。然而，15微升RPA检测仅需18分钟。此外，抗生素敏感性测试表明，高达90%的该菌菌株对美罗培南和亚胺培南耐药；检测（某基因）的15微升RPA数据显示，仅10%（n = 3）的该菌分离株未显示阳性扩增信号，其他90%（n = 27）的分离株为阳性，证实了PCR结果。
我们证明，用于检测该菌中（某基因）的新型15微升RPA检测方法比qPCR和PCR更快、更简单。它是一种有前景的替代分子诊断工具，可用于现场和即时检测中快速有效地检测该菌及耐药基因。
