摘要
利用硫氰酸胍裂解病毒和二氧化硅颗粒能够特异性吸附核酸的特性，将二氧化硅悬液、裂解液及肝炎病毒感染血清共同保温，病毒裂解释放出的核酸结合在二氧化硅颗粒上，经过双蒸水洗脱，所得核酸直接作ＰＣＲ扩增。以该法提取感染血清样本中的ＨＢＶＤＮＡ或ＨＣＶＲＮＡ作为模板，再进行ＰＣＲ扩增检测，产物在琼脂糖凝胶电泳上所见条带清晰。用本法检测ＨＢｓＡｇ（＋）／ＨＢｅＡｇ（＋）Ａｕｔｉ－ＨＢｃ（＋）血清３２例，阳性率为９６％；检测抗－ＨＣＶ阳性血清１１０例，阳性率为８２．７％。该方法成本低，实验周期短，结果稳定。
A rapid and sensitive protocol for purification of nucleic acids(DNA and RNA) from serum is reported.The method is based on the lysing property of guanidinium thiocyanate together with the nucleic acids-binding property of silica.The silica particals were mixed both with splitting buffer and HBV/HCV infected serum in 1.5ml eppendorf reaction tube at room temperature.The released nucleic acids of virions were bound to the silica particals and subsquently eluted in a low salt buffer.The purified nucleic acid was a good template for PCR.Using this procedure,32 sera being positive to HBsAg,HBeAg and anti-HBc were tested with PCR and HBV DNA was detected in 96%;110 sera being positive to anti-HCV were tested with RT-PCR and HCV RNA was picked up in 82.7%.
出处
《临床检验杂志》
CAS
CSCD
北大核心
1995年第2期68-70,共3页
Chinese Journal of Clinical Laboratory Science
关键词
核酸
多聚酶链反应
二氧化硅
肝炎病毒
Nucleic acids
Polymerase chain reaction
Silica Hepatitis B virus
Hepatitis C virus
