中国寄生虫学与寄生虫病杂志 ›› 2013, Vol. 31 ›› Issue (5): 4-346-351.

• 论著 • 上一篇    下一篇

弓形虫感染间接免疫荧光检测试剂盒的检测效果评价

卢致民1,王燕1,张子扬2,唐宏炜1,索勋3 *   

  1. 1 河北北方学院医学检验学院,张家口 075000;2 贵阳中医学院骨伤学院2011级,贵阳 550002;3 中国农业大学动物医学院,国家动物原虫实验室,北京 100193
  • 出版日期:2013-10-30 发布日期:2014-07-24

Evaluation of an Indirect Immunofluorescence Assay Kit for the Dectetion of Anti-Toxoplasma gondii IgG

LU Zhi-min1,WANG Yan1,ZHANG Zi-yang2,TANG Hong-wei1,SUO Xun3 *   

  1. 1 College of Laboratory Medicine, Hebei North University, Zhangjiakou 075000, China;2 School of Orthopaedic and Traumatology, Guiyang College of Traditional Chinese Medicine, Guizhou 550002, China; 3 College of Veterinary Medicine, China Agricultural University;National Animal Protozoa Laboratory,Beijing 100193, China
  • Online:2013-10-30 Published:2014-07-24

摘要: 目的  研制检测弓形虫IgG抗体的间接免疫荧光(IFA)检测试剂盒。  方法  在成功建立弓形虫感染IFA检测方法的基础上,构建人弓形虫感染IFA检测试剂盒。对弓形虫阳性血清、羊抗人FITC-IgG的最佳稀释浓度进行优化。对试剂盒的敏感性、特异性、稳定性和保存期等进行评价,并与国外同类试剂盒比较。  结果  弓形虫阳性血清、羊抗人FITC-IgG的最佳稀释浓度分别为1 ∶ 40和1 ∶ 100。该试剂盒可检测到弓形虫阳性血清的最大稀释倍数为1 ∶ 640,与恶性疟、间日疟、血吸虫病、棘球蚴病和囊尾蚴病患者血清均无交叉反应,但与类风湿因子阳性患者血清有交叉反应。该试剂盒的敏感性、特异性、阳性预测值、阴性预测值、符合率和Youden指数分别为90.9%、100%、100%、96.2%和97.2%和0.91,而国外同类试剂盒分别为100%、98%、95.7%、100%和98.6%和0.98,两种试剂盒的敏感性和特异性间差异无统计学意义(P>0.05)。  结论  该试剂盒检测弓形虫IgG抗体敏感性和特异性较高。

关键词: 弓形虫, 弓形虫病, 间接免疫荧光试验, IgG抗体

Abstract: Objective  To develop an indirect immunofluorescence assay (IFA) kit for detecting anti-Toxoplasma gondii IgG.  Methods  Based on the established IFA method, we established an IFA kit for the detection of human T. gondii infection. The optimal working concentrations of T. gondii IgG-positive human serum and FITC-labeled goat anti-human IgG antibody were determined. Sensitivity, specificity, reproducibility, and storage period of this kit were studied, and compared with an imported kit.  Results  The optimal working concentration of T. gondii IgG-positive human serum and FITC-labeled goat anti-human IgG antibody was 1 ∶ 40 and 1 ∶ 100, respectively. The maximum dilution of T. gondii IgG-positive human serum that the kit can detect was 1 ∶ 640. No cross reaction was observed with sera from patients with vivax malaria, falciparum malaria, schistosomosis, echinococcosis, or cysticercosis. Cross reaction was observed to the rheumatoid factor positive sera. The sensitivity, specificity, positive predictive value, negative predictive value, concordance, Youden index of this kit was 90.9%, 100%, 100%, 96.2%, 97.2%, and 0.91, respectively; and that of the imported kit was 100%, 98%, 95.7%, 100%, 98.6%, and 0.98, respectively. There was no significant difference in sensitivity and specificity between the two kits (P>0.05).  Conclusion  The IFA kit shows adequate sensitivity and specificity for detection of anti-T. gondii IgG.

Key words: Toxoplasma gondii, Toxoplasmosis, Indirect immunofluorescence assay, IgG antibody