基于PCR-化学发光的HBV/HCV/HIV并行核酸检测试剂盒的评价
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江苏省科技厅2012年第十一批省级科技创新与成果转化(生命健康科技)专项(BL2012067);江苏省卫生与计划生育委员会科技项目(H201514)


Evaluation of a parallel assay kit for detecting HBV/HCV/HIV based on PCR-chemiluminescence
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    摘要:

    目的:评价一种基于PCR与化学发光技术并行检测血液中HBV/HCV/HIV的试剂盒。方法:使用自制的磁珠及试剂提取病毒核酸,利用多重PCR、特异性探针及化学发光法检测目标病毒核酸的信息,评价试剂盒的灵敏度、特异性及稳定性等。结果:本试剂盒灵敏度高,特异性和稳定性好,能够同时检测出HBV、HCV和HIV的极限值分别为2.0、3.7和166.6 U/mL,对于可能产生干扰的10种病毒和2种细菌的病原体无交叉反应,有效期约12个月。在健康献血者10 422例酶免疫分析法合格标本中,筛查出HBV DNA核酸阳性标本12例和HCV RNA核酸阳性标本2例,与国家批准的血液筛查核酸试剂平行检测结果一致。结论:该试剂盒快速、准确、高灵敏、低成本,适用于基因诊断及流行病学调查。

    Abstract:

    Objective: To evaluate a HBV/HCV/HIV nucleic acids parallel assay kit via PCR and chemiluminescence. Methods: The viral nucleic acids of HBV, HCV and HIV were simultaneously extracted with the homemade magnetic nanoparticles and reagents. The information on the target viral nucleic acids was checked by multiple PCR, electrophoresis, and chemiluminescence. The sensitivity, specificity and stability of the kit were also evaluated. Results: The kit was highly sensitive, specific and stable. The detection limits of the kit were 2.0, 3.7 and 166.6 U/mL for HBV DNA, HCV RNA and HIV RNA, respectively. There was no cross-reactivity with the 10 kinds of viruses, and 2 kinds of bacteria, which might cause interference potentially. The expiry date was about 1 year. Among the 10 422 EIA negative samples of blood donors, 12 were HBV positive and 2 were HCV positive. These results completely coincided with those of other current diagnostic methods of commercial kit applied in blood screening. Conclusion: The kit is fast, accurate, highly sensitive and cost-effective. It can be applicable to the genetic diagnosis and epidemiological survey.

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张若洋,范恩勇,王明元,Zeeshan Ali,汪久海,冯晨晨,吴敏慧,史丽莉,李智洋,梁文飚.基于PCR-化学发光的HBV/HCV/HIV并行核酸检测试剂盒的评价[J].南京医科大学学报(自然科学版),2017,(3):335-339

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  • 收稿日期:2016-12-27
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  • 在线发布日期: 2017-04-07
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