摘要
同一份血清中的ＨＢＶＤＮＡ与ＨＣＶＲＮＡ经热变性直接法一步裂解，先用针对ＨＣＶ特异的外引物将ＨＣＶＲＮＡ逆转录为ｃＤＮＡ，然后采用ＨＢＶ，ＨＣＶ各自特异的２套引物进行ＰＣＲ同步扩增。结果按预定大小，ＰＣＲ产物出现２条带。分别为４２８ｂｐ，１４４ｂｐ。将产物转移至尼龙膜上，经α－３２ＰｄＣＴＰ标记ＨＣＶ探针及地高辛素标记ＨＢＶ探针进行重复杂交，证明１４４ｂｐ为ＨＣＶ所特有，４２８ｂｐ为ＨＢＶ特有。用该方法对３０份单独检测证实ＨＢＶ，ＨＣＶ核酸均阳性血清测验，其结果完全吻合、该二联技术可明显缩短检测时间，具有简便、快速、敏感的特点。
new technique which amplified HBV DNA and HCV RNA in serum at the same time was established. The templat was extracted by a hot-denatured method. Reverse transcrip- tion was carried out, used primer derived from HCV genome alone , then HBV DNA and HCV RNA was amplified simultaneously by HBV and HCV nested primers respectively. The preassigned size of amplified products was 428 bp and 144 bp , which were determined in ethi- olium bromide (EB) stained agarose gel after electroresis under ultraviolet (UV) light. The products were electrophoretically transfered to nylon membrance and rehybriated with α-32P and digoxisine-labelled probes of HCV and HBV respectively.
出处
《中山医科大学学报》
CSCD
1996年第2期145-148,共4页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
广东省科委资助课题
关键词
乙肝病毒
丙肝病毒
聚合酶链反应
DNA
RNA
DNA, viral/analysis
RNA , viral /analysis
hepatitis B virus
hep- atitis C virus
polymerase chain reaction
