摘要
目的 建立用于 SEN病毒 (SENV)检测的巢式聚合酶链反应 (n PCR)方法。 方法 选择 SENV各亚型间高度保守的 5′端非编码区 (5′-NCR)序列设计 2对特异性引物 ,建立可同时检测 SENV所有亚型的 n PCR方法。对 1 73份慢性乙型肝炎 (CHB)患者血清和 96份正常人血清进行 SENV检测 ,并随机选择 3份 PCR阳性产物进行克隆测序。 结果 倍比稀释实验表明该方法的终检稀释度为 1 0 3。PCR产物克隆测序结果显示 :各分离株与 Genbank收录的 SENV序列相应区段同源性为 95 %～ 96%。CHB患者 SENV感染率为 86.1 % ,正常人为 85 .1 % ,二者差异无显著性 (χ2 =0 .3 81 ,P=0 .5 3 7)。 结论 该方法敏感、特异且简便经济 ,适于开展
Objective\ To establish a nested polymerase chain reaction (nPCR) method for detecting SENV. Methods\ Two pairs of primers were designed from highly conservative sequence of SENV A～H on the 5′ terminal non coding region.The nested PCR method for SENV A～H was built and the specificity and sensitivity were tested.Sera of 173 chronic hepatitis B patients (CHB) and 96 normal individuals were detected by this method.\ Results\ The maximum dilution of serum samples is 10 3 using this method for detection of SENV.Sequence analysis showed 95%～96% nucleotides of cloned SENV strains were analogous to the SENV genome registered in Genbank.The prevalence rates of SENV were 86 1% in CHB patients and 85 1% in normal individuals respectively (χ 2=0 381, P =0 537).\ Conclusions\ This method of nested PCR is sensitive,reliable and economical,and can be applied to the epidemiological survey of SENV D/SENV H.
出处
《临床输血与检验》
CAS
2002年第3期15-17,共3页
Journal of Clinical Transfusion and Laboratory Medicine
基金
北京市委组织部优秀人才项目资助 (2 0 0 1-0 1)
