聚合酶链反应检测血清丁型肝炎病毒核酸

Reverse transcription-"nested"polymerase chain reaction for detection of HDV RNA in serum sam-ples.

目的为建立一种特异性强、敏感性高的检测丁型肝炎病毒(HDV)感染的方法,提高丁型肝炎的诊断水平。方法根据 HDV RNA 保守区(654～963位核苷酸)设计合成两对引物,采用逆转录一套式聚合酶链反应(RT-"nested"PCR)对35例肝炎患者血清进行检测。结果二次 PCR 扩增的特异性和敏感性均较一次 PCR 高。19例丁型肝炎抗原阳性者 HDV RNA 均为阳性,9例丁型肝炎抗体阳性者中有6例阳性,7例单纯 HBsAg 和 HBeAg 阳性者均未检出 HDV RNA。结论这一检测方法特异性强、敏感性高,可广泛用于临床检测。

Objective It is essential to provide a highly sensitive and specific method for the detec- tion of hepatitis D virus(HDV)to improve the diagnosis of HDV infection.Methods The reverse transcription-'nested'polymerase chain reaction(RT-'nested'PCR)method was developed to directly detect HDV RNA in serum samples.RT-'nested' PCR was carried out by using two pairs of primers based on the conserved region(654～963 nucleotides)of the HDV RNA sequence in 35 serum samples from hepatitis patients;Results HDV RNA detection by 'nested'PCR was more sensitive and specific than by one time PCR.HDV RNA was detected in 19/19 of the serum samples positive for HDAg and 6/9 of the serum samples positive for anti-HD,while HDV RNA was negative in 7 HBsAg and HBeAg serum samples.Conclusions The RT-'nested'PCR described is a very specific and sensitive method which may be used in routine detection of HDV RNA.