同时检测SEN病毒D和H聚合酶链反应法的建立  

Simultaneous detection of SENV-D and SENV-H with polymerase chain reaction

作　　者：董庆鸣[1] 何忠平[1] 庄辉[2] 闫杰[1] 宋淑静[1] 王小红[2] 

机构地区：[1]北京地坛医院,100011 [2]北京大学医学部病原生物学系

出　　处：《中国公共卫生》2003年第1期39-41,共3页Chinese Journal of Public Health

基　　金：北京市卫生局肝炎重点学科基金项目

摘　　要：目的　建立同时检测SEN病毒D和H(SENV -D和H)巢式聚合酶链反应法 (nPCR)。方法　第一轮PCR外引物为SENV -D和H开放读码框架 (ORF) 1区的共同序列 ,第二轮PCR内引物分别为SENV -D和HORF1区型特异性引物。应用双脱氧链末端终止法对SENV -D和H的PCR产物进行克隆测序。结果　应用同时检测SENV -D和H的nPCR法最终检出SENV -D和H的血清稀释度为 10 3。序列分析表明 ,用本法检出的SENV -D(DTd株 )和SENV-H(DTh株 )与GenBank收录的SENV -D和H株核苷酸序列同源性均为 92 %。检测 173例慢性乙型肝炎 (轻、中度 )患者 ,SENV -D和H的总感染率为 5 8 4%。结论　本法可用于SENVObjective To establish a nested polymerase chain reaction (nPCR) for simultaneous detection of SENV-D and H.Methods The outer primers of the first-round PCR were derived from the conserved region of ORF1 of SENV-D and H.Two sets of specific inner primers of the second-round PCR were obtained from ORF1 of SENV-D and SENV-H,respectively.The products of the nPCR were cloned into plasmids and sequenced by the dideoxy-mediated chain-termination method.Results The maximum dilution of serum was 10\+3 for detection of SENV-D and H using the nPCR.Sequence analysis showed the nucleotide homology was 92% between the cloned SENV strains (DTd and DTh) and SENV-D and SENV-H registered in GenBank,respectively.The total rate of SENV-D and H infections was 58.4% in patients with chronic hepatitis B.Conclusion This method can be used for the clinical diagnosis and epidemiological study of SENV-D and H infections.

关 键 词：SEN病毒 巢式PCR 乙型肝炎 流行病学 临床诊断 

分 类 号：R512.62[医药卫生—内科学]