中国寄生虫学与寄生虫病杂志

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基于捕获与连接的探针式PCR矩阵拆分混样检测疟原虫的效果评价

周耀武1,林祖锐1,程志斌2,郑直2,周兴武1,周子悠1,王多全3,李建雄1,曾旭灿1,孙晓东1*   

  1. 1云南省寄生虫病防治所,云南省疟疾研究中心,云南省虫媒传染病防控研究重点实验室,普洱 665000;2中国医学科学院基础医学研究所,北京 100005;3中国疾病预防控制中心寄生虫病预防控制所,世界卫生组织热带病合作中心,科技部国家级热带病国际联合研究中心,卫生部寄生虫病原与媒介生物学重点实验室,上海 200025
  • 出版日期:2016-08-30 发布日期:2016-11-07
  • 基金资助:

    云南省中青年学术技术带头人后备人才项目(No. 2015HB075)

Efficacy of Capture and Ligation Probe-PCR with#br# Pooling Strategy in Detection of Plasmodium spp.

ZHOU Yao-wu1, LIN Zu-rui1, CHENG Zhi-bin2, ZHENG Zhi2,ZHOU Xing-wu1, ZHOU Zi-you1, WANG Duo-quan3, LI Jian-xiong1, ZENG Xu-can1, SUN Xiao-dong1*   

  1. 1 Yunnan Institute of Parasitic Disease, Yunnan Center for Malaria Research, Yunnan Key Laboratory of Vector-borne Diseases Control and Research, Pu’er 665000, China;2 Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Beijing 100005, China; 3 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention;WHO Collaborating Centre for Tropical Diseases;National Center for International Research on Tropical Diseases, Ministry of Science and Technology;Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China
  • Online:2016-08-30 Published:2016-11-07
  • Supported by:

    Supported by the Candidates of the Young and Middle Aged Academic Leaders of Yunnan Province(No. 2015HB075)

摘要:

采用基于捕获与连接的探针式PCR(capture and ligation probe-PCR,CLIP-PCR)矩阵拆分混样法和镜检两种方法对100份临床样本进行检测,综合评价CLIP-PCR矩阵拆分混样法对疟原虫的检测效果。从云南省疟疾参比实验室标本库选取93份阴性样本和7份阳性样本,随机混合,采用双盲法进行CLIP-PCR矩阵拆分混样检测,并与镜检法的结果对比分析,通过检出率、灵敏度、特异度、检测时间及检测成本等进行综合评价。7份镜检阳性样本被全部检出,检出率为100%。以镜检为金标准,CLIP-PCR矩阵拆分混样检测灵敏度和特异度均为100%。全部样本的检测时间为5.0 h,较镜检的33.3 h缩短85.0%;检测成本为300元,较镜检的1 000元降低了70.0%。

关键词: 捕获与连接的探针式-PCR, 矩阵拆分混样, 效果评价

Abstract:

The capture and ligation probe-PCR(CLIP-PCR) with pooling strategy method and microscopy were applied on 100 clinical samples(7 positive and 93 negative samples) from the malaria reference laboratory in Yunnan Province. By calculating the detection rate, sensitivity, specificity, detection time and detection cost, the efficacy of the CLIP-PCR with pooling strategy method in detecting Plasmodium spp. was evaluated. The CLIP-PCR with matrix pooling strategy successfully detected Plasmodium spp. in all the 7 positive samples. Its sensitivity and specificity relative to the microscopy as a gold standard were both 100%. The detection time for all the samples by CLIP-PCR was 5.0 h, 85.0% shorter than that by microscopy(33.3 h), and the detection cost was 300 yuan, 75.0% less than that by microscopy (1 000 yuan).

Key words: Capture and ligation probe-PCR, Pooling strategy, Evaluation