恶性疟与间日疟双标记时间分辨免疫荧光检测试剂盒的研制

发布时间：2018-01-16 23:05

本文关键词：恶性疟与间日疟双标记时间分辨免疫荧光检测试剂盒的研制　出处：《中国病原生物学杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的采用双标记时间分辨荧光免疫分析(TRFIA)技术研制一种同时检测恶性疟原虫、间日疟原虫及混合感染试剂盒,并评价试剂盒各项性能指标。方法用抗恶性疟原虫/间日疟原虫乳酸脱氢酶(PLDH)抗体1H12作为捕获抗体包被96孔板,以Sm3+标记抗恶性疟原虫乳酸脱氢酶(PfLDH)抗体2A5和Eu3+标记抗间日疟原虫乳酸脱氢酶(PvLDH)抗体4F6作为检测抗体,建立恶性疟与间日疟双标记时间分辨荧光分析法。结果试剂盒检测恶性疟原虫的灵敏度为0.15ng/ml,线性范围为0.5500ng/ml,平均稀释回收率为102.2%,批内与批间变异系数分别为6.48%和6.63%;检测间日疟原虫的灵敏度为0.25ng/ml,线性范围为0.5500ng/ml,平均稀释回收率为100.78%,批内与批间变异系数分别为5.39%和6.16%,恶性疟与间日疟检测不相互干扰。用自制试剂盒检测212份疟疾患者血标本,阳性率为95.28%,高于镜检法的92.45%和Optimal法的51.89%。结论 PfLDH/PvLDH-TRFIA试剂盒能同时检测恶性疟、间日疟及二者混合感染,且灵敏度高,可测范围宽,稳定性好,具有良好应用前景。
[Abstract]:Objective to develop a kit for simultaneous detection of Plasmodium falciparum, Plasmodium vivax and co-infection by double labeling time-resolved fluorescence immunoassay (TRFIA). Methods the anti-Plasmodium falciparum / Plasmodium vivax lactate dehydrogenase (PLDDH) antibody 1H12 was used as the capture antibody coated with 96 well plate. Sm3 labeled anti-Plasmodium falciparum lactate dehydrogenase antibody 2A5 and Eu3 labeled anti-Plasmodium vivax lactate dehydrogenase antibody 4F6 were used as detection antibodies. A double-label time-resolved fluorescence assay for falciparum malaria and vivax malaria was established. Results the sensitivity of the kit for the detection of Plasmodium falciparum was 0.15 ng / ml, and the linear range was 0.5500 ng / ml. The average dilution recovery was 102.2, and the coefficient of variation was 6.48% and 6.63, respectively. The sensitivity of detection of Plasmodium vivax was 0.25 ng / ml, the linear range was 0.5500 ng / ml, and the average dilution recovery was 100.78%. The coefficients of variation within and between batches were 5.39% and 6.16, respectively. Falciparum malaria and vivax malaria were not interfered with each other. A self-made kit was used to detect 212 blood samples of malaria patients, and the positive rate was 95.28%. It is higher than that of Optimal and 92.45%. Conclusion PfLDH/PvLDH-TRFIA kit can be used to detect falciparum malaria at the same time. Vivax malaria and its mixed infection have high sensitivity, wide range of detection, good stability, and have a good prospect of application.
【作者单位】：山东省医学科学院山东省寄生虫病防治研究所山东省医学科学院第三附属医院;山东医药技师学院;广州优迪生物科技有限公司;解放军第88医院;
【基金】：广东省科技型中小企业技术创新专项资金项目(No.2016A010119067)
【分类号】：R531.3
【正文快照】： ***疟疾是由疟原虫感染引起的虫媒传染病,寄生于人类的疟原虫有恶性疟原虫、间日疟原虫、三日疟原虫和卵形疟原虫4种,其中恶性疟原虫和间日疟原虫危害最大。据WHO报道,近年全球范围内有2.14亿起***疟疾病例,43.8万人死亡,约32亿人面临疟疾风险[1]。快速准确地诊断早期疟原虫感

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