中国媒介生物学及控制杂志 ›› 2010, Vol. 21 ›› Issue (1): 39-42.

• 论著 • 上一篇    下一篇

4种鼠疫F1抗原/抗体检测方法的初步评价

王艳华,海荣,夏连续,蔡虹,梁莹,俞东征   

  1. 中国疾病预防控制中心传染病预防控制所人兽共患病室,传染病预防控制国家重点实验室(北京102206)
  • 收稿日期:2009-08-27 出版日期:2010-03-01 发布日期:2010-03-01
  • 通讯作者: 俞东征,Email: yudongzheng@icdc.cn
  • 作者简介:王艳华(1971-),女,博士,副研究员,从事人兽共患病的病原学及分子生物学研究。

Preliminary evaluation of four methods to determine the F1 antigen/antibody of plague

 WANG Yan-Hua, HAI Rong, XIA Lian-Xu, CAI Hong, LIANG Ying, YU Dong-Zheng   

  1. Department of Zoonosis, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention; State Key Laboratory for Infectious Disease Prevention and Control, Beijing 102206, China
  • Received:2009-08-27 Online:2010-03-01 Published:2010-03-01
  • Contact: YU Dong-zheng, Email: yudongzheng@icdc.cn

摘要:

目的 客观评价血凝试验、胶体金试验、ELISA和PCR 4种检测方法的灵敏性。方法 对于血清学方法中的F1抗体检测,评价检出的全菌免疫兔抗血清最低稀释度;F1抗原检测,评价检出的最低浓度。对于分子生物学方法,评价同时检出fra基因和pla基因的模板最低浓度。结果 对于F1抗体检测,间接血凝试验检出的最低稀释度是1∶64,胶体金试验是1∶1000,ELISA是1∶204 800。对于F1抗原检测,反向血凝试验检出的最低浓度是2 ng/ml,胶体金试验是50 ng/ml。fra、 pla基因同时检出,模板的最低浓度是0.21 ng/μl。结论 在以抗体IgG为主的血清中,ELISA法对于F1抗体检测的灵敏性较高。反向间接血凝试验对于F1抗原检测的灵敏性较高。为减少误诊率,采用PCR诊断鼠疫菌要求最低模板浓度是0.21 ng/μl。

关键词: 鼠疫耶尔森菌, 血凝试验, 胶体金试验, 酶联免疫吸附试验, 聚合酶链反应

Abstract:

Objective To evaluate the sensitivity of the four test methods, hemagglutination test, gold immunochromatographic assay (GICA),  enzyme-linked  immune  sorbent assay (ELISA)  and  polymerase  chain  reaction (PCR).  Methods The  minimal detectable dilution of antiserum of a bacteria?immunity rabbit via serological approaches for F1 antibody detection, the minimal detectable concentration in F1 antigen measurement, and the minimal concentration of template with a detectable gene pair, fra and pla, were evaluated. Results For the F1 antibody test, the minimal dilution detected was 1∶64 through IHA, 1∶1000 through GICA, and 1∶204 800 through ELISA. For F1 antigen test, the minimal detectable concentration was 2 ng/ml by RIHA and 50 ng/ml by GICA. The minimal concentration of template was 0.21 ng/μl when fra and pla were both detected. Conclusion In the serum consisting mainly of antibody IgG, the ELISA method has a higher sensitivity detecting F1 antibodies. The RIHA has higher sensitivity detecting F1 antigens. To minimize misdiagnosis, a minimal concentration of template of 0.21 ng/μl is required to diagnose plague when using PCR.

Key words: Yersinia pestis, Hemagglutination test, Gold immunochromatographic assay, Enzyme-linked immune sorbent assay, Polymerase chain reaction

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