华南预防医学 ›› 2015, Vol. 41 ›› Issue (6): 307-311.doi: 10.13217/j.scjpm.2015.0307

• 中东呼吸综合征防控 •    下一篇

中国首例中东呼吸综合征病例的实验室检测与商品化诊断试剂盒的评价

邹丽容1,徐静1,2,武婕1,宋颖超1,梁丽君1,张欣1,柯昌文1,2   

  1. 1.广东省疾病预防控制中心,广东省新发传染病病原学检测技术研究中心,广东省病原微生物菌毒种保藏和利用医学重点实验室,广东 广州 511430;2.南方医科大学
  • 出版日期:2015-08-20 发布日期:2015-10-12
  • 通讯作者: 柯昌文 E-mail:kecw1965@aliyun.com
  • 作者简介:邹丽容(1975—),女,硕士研究生,副主任技师,研究方向为呼吸道病毒的监测与研究;徐静(1989—),女,在读硕士研究生,研究方向为传染病病原体快速分子诊断方法的建立与评估

Laboratory diagnosis of the first imported case of Middle East respiratory syndrome in China and evaluation of commercial kits for MERS-CoV

ZOU Li-rong1,XU Jing1,2,WU Jie1,SONG Ying-chao1,LIANG Li-jun1,ZHANG Xin1,KE Chang-wen1,2   

  1. 1.Key Laboratory for Repository and Application of Pathogenic Microbiology, Research Center for Pathogens Detection Technology of Emerging Infectious Diseases, Guangdong Provincial Center for Disease Control and Prevention, Guangzhou 511430, China;2.Southern Medical University
  • Online:2015-08-20 Published:2015-10-12

摘要: 目的对广东省首例中东呼吸综合征(MERS)病例进行实验室快速诊断,并比较2种不同品牌商品化试剂盒对中东呼吸综合征冠状病毒(MERS-Co V)的检测效果。方法于2015年5月28日采集首例韩国输入疑似MERS病例的3份咽试子和1份血清样本,以及陆续采集50份密切接触者的咽试子样本,根据WHO和国家MERS实验室检测技术指南推荐的荧光定量PCR方法,分别使用针对up E、ORF1a、ORF1b 3个目的基因的3对引物探针开展检测。利用首例MERS病例的阳性核酸提取物分别对A、B 2种商品化MERS-Co V RNA荧光定量PCR检测试剂盒的灵敏度和特异度进行评估。结果疑似MERS病例的3份咽拭子样本同时检出up E、ORF1a、ORF1b 3个目的基因和1份血清样本同时检出up E、ORF1a 2个目的基因,均证实为MERS-Co V核酸阳性。A、B 2种试剂盒检测病例的3份咽拭子样本均为MERS-Co V核酸阳性,并能检测到经10倍稀释的MERS-Co V核酸提取物,但均未能检测出病例血清样本阳性。使用国家推荐荧光定量PCR方法及A、B 2种商品化试剂盒检测50份病例密切接触者的咽拭子样本的MERS-Co V核酸,结果均为阴性。结论首次对1例输入性MERS病例进行了实验室快速诊断;WHO和国家MERS实验室诊断技术指南中的荧光定量PCR的病毒检测方法,以及A、B 2种商品化试剂盒均可用于MERS-Co V感染的实验室诊断。

Abstract: ObjectiveTo conduct rapid laboratory diagnosis of the first imported case of the Middle East respiratory syndrome coronavirus (MERS-CoV) infection in Guangdong Province and compare the testing performance of two commercial kits for MERS-CoV.MethodsThree pharyngeal swabs and 1 serum sample were collected from the first suspected MERS-CoV infection case imported from the South Korea on May 28, 2015.And 50 pharyngeal swabs were collected from close contacts in succession.According to the guidelines for laboratory testing for MERS-CoV by WHO and China Center for Disease Control and Prevention (CCDC), Guangdong Provincial Center for Disease Control and Prevention (GDCDC) had prepared three sets of primers and probes for MERS-CoV upE, ORF1a, and ORF1b genes to carry out Real time RT-PCR.The sensitivity and specificity of two commercial MERS-CoV RNA fluorescence quantitative PCR detection kits A and B were evaluated by using the positive nucleic acid extracts of the first MERS case, respectively.ResultsThe target genes of upE, ORF1a, and ORF1b were detectable in 3 throat swabs.and upE and ORF1a, in the serum sample of the suspected MERS case.Therefore, the MERS-CoV nucleic acid was confirmed to be positive.The MERS-CoV was found to be positive in the 3 throat swab samples of the case detected by both commercial kits A and B, which can also detected the 10 times diluted extracted nucleic acid of MERS-CoV.All 50 samples from the close contacts with the case were MERS-CoV nucleic acid negative detected by the fluorescence quantitative PCR recommended by CCDC and the two kits of A and B.ConclusionThe first imported suspected MERS case was positive for MERS-CoV by rapid laboratory diagnosis.The guidelines for laboratory testing for MERS-CoV and the two commercial kits can be used as laboratory diagnosis tools of MERS-CoV infection.

中图分类号: 

  • R183.3