适用于中国人群的隐匿性乙肝病毒感染的检测方法优化研究

发布时间：2018-04-05 18:34

本文选题：乙肝病毒　切入点：隐匿性感染　出处：《基因组学与应用生物学》2017年10期

【摘要】：隐匿性乙肝病毒感染的诊断,通常使用巢式PCR的方法,来检测血清中的HBV DNA。因中国人群感染乙肝病毒的基因型与国外人群存在差异,故本研究的目的是对现有文献已经介绍的诊断隐匿性乙肝病毒感染的PCR检测方法进行优化,从而适用于中国隐匿性乙肝病毒感染人群的诊断。本研究通过对文献报道的巢式PCR检测方法进行验证,发现其Core/pre-core区段扩增结果不理想,故根据我国乙肝病毒的基因型以及HBV C基因的保守性原则自行设计优化实验。利用临床收集的HBV阳性样本验证其灵敏度及特异度;收集父亲或/和母亲为HBV阳性患者,而孩子HBs Ag结果为阴性的样本来验证其扩增结果。本研究优化设计的检测HBV C基因的引物,具有灵敏度高特异性好的优点。本研究临床收集了100份父亲或/和母亲为HBV阳性患者,而孩子HBs Ag结果为阴性的血清样本,用本研究优化后的引物检测,可以提高隐匿性乙肝病毒的检测分辨率,提高了4倍的检测效率。本研究优化设计的隐匿性乙肝病毒感染的引物可以用于中国人群中隐匿性乙肝病毒感染的检测,具有较高的灵敏度和特异性。
[Abstract]:In the diagnosis of occult hepatitis B virus infection, nested PCR is usually used to detect HBV DNA in serum.Because the genotype of hepatitis B virus infection in Chinese population is different from that in foreign population, the purpose of this study is to optimize the PCR detection method for the diagnosis of occult hepatitis B virus infection, which has been introduced in the existing literature.Thus it is suitable for the diagnosis of occult hepatitis B virus infection in China.This study verified the nested PCR detection method reported in literature and found that the amplification results of Core/pre-core region were not satisfactory. Therefore, the optimal experiment was designed according to the genotype of hepatitis B virus and the conservative principle of HBV C gene in China.The sensitivity and specificity of HBV positive samples were verified by clinical samples, and the amplification results were verified by collecting samples whose father or / and mother were HBV positive and children were HBs Ag negative.The optimized primer for detecting HBV C gene has the advantages of high sensitivity and good specificity.In this study, we collected 100 serum samples whose father or / and mother were HBV positive and their children were HBs Ag negative. The detection resolution of occult hepatitis B virus could be improved by using the optimized primers.The detection efficiency is increased by 4 times.The primers optimized in this study can be used for the detection of occult hepatitis B virus infection in Chinese population with high sensitivity and specificity.
【作者单位】：重庆医科大学附属儿童医院儿科研究所儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地儿童感染免疫重庆市重点实验室;十堰市太和医院(湖北医药学院附属医院)儿童医疗中心PICU;贵州省人民医院儿科;
【基金】：国家自然科学基金(81371876) 重庆市卫生局重点项目(2013-39-1-025) 重庆市高等学校优秀人才资助计划(2014-47-1-11)共同资助
【分类号】：R512.62

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