聚合酶链反应检测慢性丁型肝炎血清HDV RNA的实验研究和临床观察

DETECTION OF SERUM HDV RNA BY POLYMERASE CHAIN REACTION IN PATIENTS WITH HEPATITIS DELTA

本文建立一种高度敏感和特异性的检测丁型肝炎病毒(HDV)RNA的方法,提高了丁型肝炎的诊断水平。以HDV RNA保守区ORF5′末端第929～1640位核苷酸为靶基因设计一对引物,采用逆转录-聚合酶链反应(RT-PCR)检测了35例慢性丁型肝炎患者血清HDV RNA,并同时检测HBVM。35例中共检出HDV RNA421例(60％),10例HDAg阳性者HDV RNA全部阳性,25例抗-HD阳性者中有11例HDV RNA阳性(44％)。表明采用RT-PCR可以准确、快速、敏感地检测出HDV RNA,具有很强的特异性。35例中HBV DNA的检出率为34.2％,而HDV RNA的检出率为60％,HBV DNA和HDV RNA同时阳性者共9例占25.7％,提示慢性乙型肝炎病人合并丁型肝炎时,HBV的复制受到不同程度的抑制而HDV的复制较为活跃。

The reverse transcription polymerase chain reaction (RT-PCR) was carried out using a pairs of primmers based on the conserved region at the 5 ends(929-1640) nucleotides of the HDV RNA sequence in 35 serum samples from chronic hepatitis D.HDV RNA was positive in 100% of the patients with HDAg positive and in 44% of with anti-HD positive.The results of this study show that HDV RNA determination by RT-PCR assay is a very specific and sensitive method for the diagnosis of HDV infection.HBV DNA was positive by PCR in 12(34.28%) cases and HDV RNA was positive in 21 (60% ) cases of the 35 patients with chronic hepatitis D.9(25.71%) was found of the two viral nucleic acids positive simultaneously.our study has shed light on support the phenomenon that HBV replication is suppressed in chronic hepatitis B patients with HDV coinfection.