| 李志兰,蒋秀娣,陈佩宏,别立翰,郭乃华.应用环介导等温扩增技术检测铜绿假单胞菌OprD2基因的临床研究[J].,2018,(17):3274-3279 |
| 应用环介导等温扩增技术检测铜绿假单胞菌OprD2基因的临床研究 |
| Clinical Study on Detection of Pseudomonas Aeruginosa OprD2 Gene by Loop Mediated Isothermal Amplification |
| 投稿时间：2017-11-26 修订日期：2017-12-21 |
| DOI：10.13241/j.cnki.pmb.2018.17.015 |
| 中文关键词: 铜绿假单胞菌 OprD2基因 Real-time PCR LAMP 应用价值 |
| 英文关键词: Pseudomoas aeruginosa OprD2 gene Real-time PCR LAMP Application value |
| 基金项目:上海市浦东新区卫生和计划生育委员会卫生科技资助项目（PW2014B-13） |
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| 中文摘要: |
| 摘要 目的：评价检测铜绿假单胞菌外膜孔蛋白OprD2基因缺失的环介导等温扩增技术（LAMP）的应用价值，从而为临床早期诊断耐亚胺培南铜绿假单胞菌（IRPA)以及治疗方案的制定提供参考依据。方法：收集2016年1月至2016年12月上海中医药大学附属第七人民医院临床分离的IRPA，MicroScan WalkAway-96 plus全自动鉴定及药敏系统检测最低抑菌浓度(MIC)，应用LAMP法检测OprD2基因的缺失，并与实时荧光定量PCR（Real-time PCR）法进行比较。结果：50株样本中，DNA A260/280均≧1.72，而DNA浓度均>370 ng/μL，6号和49号样本甚至>2000 ng/μL。LAMP法检测50株IRPA的OprD2 基因，结果显示26株阳性，其余24株为阴性，缺失率为48.0%；Real-time PCR法检测结果显示，50株IRPA的OprD2 基因阳性和阴性分别为26株、24株，LAMP法检测结果与Real-time PCR法具有一致性。结论：针对靶点OprD2 基因设计引物的LAMP法是一种快速、简便且便于临床实验室应用的方法，其检测结果与Real-time PCR法具有高度一致性。 |
| 英文摘要: |
| ABSTRACT Objective: To evaluate the application of loop mediated isothermal amplification (LAMP) technique for the detection of the deletion of outer membrane protein OprD2 gene in Pseudomonas aeruginosa, it can provide reference for the early clinical diagno- sis of imipenem resistant Pseudomonas aeruginosa(IRPA) and the treatment plan. Methods: The IRPA of clinically isolated were collected from the Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine from January 2016 to December 2016, the MicroScan WalkAway-96 plus full automatic identification and drug sensitive system were used to detect the minimum inhibitory con- centration (MIC), the deletion of OprD2 gene was detected by LAMP, and compared with real-time quantitative PCR (Real-time, PCR) method. Results: Among the 50 samples, both DNA A260/280≧1.72, both DNA concentration>370 ng/μL, and even the concentration of samples 6 and 49>2000 ng/μL. The OprD2 gene of 50 strains of IRPA was detected by LAMP, the results showed that 26 strains were positive and the other 24 were negative, and the deletion rate was 48%. The results of Real-time PCR showed that the positive and nega- tive OprD2 genes of 50 strains of IRPA were 26 and 24 respectively, the result of LAMP method was consistent with that of Real-time PCR. Conclusion: LAMP method is a rapid, simple and convenient method for clinical laboratory application for the target of OprD2 gene design, and the detection result is highly consistent with the Real-time PCR method. |
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