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。
Occult HBV infection is usually diagnosed by the nest PCR method, which is used to detect HBV DNA in serum. Due to the difference of HBV genotype between Chinese and foreigners, the aim of this survey was to optimize the PCR testing methods for the diagnosis of occult HBV infection in the existing literature, so as to be suitable for the diagnosis of Chinese patients with occult HBV infection. By verifying the reported nest PCR methods, we found that the primers of the Core/pre-core did not work well in Chinese population. Therefore,according to the genotype of HBV and the conservation principle of HBV C genes, the optimization experiments were designed. Then the sensitivity and specificity of the method was verified by HBV positive samples that were collected clinically. Samples with fathers or/and mothers as HBV positive patients, while HBs Ag results of children were negative were collected to validate its amplification results. In this study, serum samples of 100 patients whose fathers and/or mothers were HBV positive patients, while the HBs Ag results of children were negative were collected. The optimized primers could improve the detection resolution of occult HBV, and the detection efficiency was improved by 4 times. The optimized primers designed for occult HBV infection could be used to detect occult HBV infection in Chinese population with high sensitivity and specificity
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2017年第10期4036-4042,共7页
Genomics and Applied Biology
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
