同时检测SEN病毒D和H聚合酶链反应法的建立

董庆鸣; 何忠平; 庄辉; 闫杰; 宋淑静; 王小红

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

摘要

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10³。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584%。
结论本法可用于SENV-D和H感染的临床诊断和流行病学调查。

Abstract:
Objective 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 10³ 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.

HTML全文

参考文献(5)

施引文献

资源附件(0)