摘要
目的建立环介导的等温扩增技术(LAMP)对HBV、HCV和H IV核酸检测的方法并对检测灵敏度和特异性作初步考核。方法通过引物设计和筛选、内质控的设计与时间分辨浊度检测方法的运用,建立LAMP HBVDNA、HCV RNA和H IV RNA扩增体系;用连续稀释的阳性样本和不含任何病毒核酸的正常人血浆考核所建立的LAMP扩增体系的灵敏度和特异性。结果建立了LAMP HBV DNA扩增体系及HCV/H IV RNA双联检体系,该体系对5 CP/m l的HBV DNA样本的检出率为51.85%,对100 CP/m l的HCV RNA阳性样本的检出率为61.90%,对100 CP/m l的H IV RNA阳性样本的检出率为45%。对考核样本检测的相对灵敏度和特异性均为100%。结论LAMP HBV、HCV和H IV检测灵敏度较高,在进一步优化以后能用于HBV、HCV和H IV的核酸检测。
Objective To establish nucleic acid testing metthods for hepatitis B virus （ HBV ）, hepatitis C virus （HCV） and human immunodeficiency virus （HIV） based on loop-mediated isothermal amplification （ LAPM） technique, and to assess the sensitivity and specificity of these tests preliminarily. Methods A series of primer sets were designed and screened. The internal control system was designed and the time discriminated turbidity detection method was established and applied. After establishing the amplification system ,serially diluted positive samples and normal person samples without any viral nucleic acid were used to evaluate the sensitivity and specificity of tbe test. Results LAMP based HBV DNA testing and HCV/HIV multiplex testing system were established. The detection rates of the tests were 51.85% （ 14/27 ）, 61.90% （ 13/21 ） and 45% （9/20 ）, respectively, when we tested 5cp/ml HBV DNA positive samples, 100cp/ml HCV RNA positive samples and 100cp/ml HIV RNA positive samples. The relative sensitivity （ 27/27） and specificity （ 30/30 ） of the test were both 100% when they were assessed with the positive and negative plasma samples. Conclusion The established LAMP nucleic acid testing methods is highly sensitive. This nucleic acid testing method could be used for HBV, HCV and HIV testing after optimization.
出处
《中国输血杂志》
CAS
CSCD
2008年第11期829-833,共5页
Chinese Journal of Blood Transfusion
基金
上海市医学领先专业重点学科建设项目--安全输血第三期资助课题(编号:05Ⅲ003)
关键词
环介导的等温扩增
核酸检测
乙型肝炎病毒
丙型肝炎病毒
人类免疫缺陷病毒
Loop-mediated isothermal amplification
Nucleic acid testing
Hepatitis B virus （HBV ）
Hepatitis C virus （ HCV ）
Human immunodeficiency virus （ HIV ）
