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DNA:用于诊断乙型肝炎感染的DNA杂交方法。

DNA: DNA hybridization method for the diagnosis of hepatitis B infection.

作者信息

Feinman S V, Berris B, Guha A, Sooknanan R, Bradley D W, Bond W W, Maynard J E

出版信息

J Virol Methods. 1984 May;8(3):199-206. doi: 10.1016/0166-0934(84)90014-4.

Abstract

Hepatitis B viral (HBV) DNA was detected in a hepatoma cell line which produces hepatitis B surface antigen (HBsAg) and in patients with acute hepatitis B. The serum of one patient with acute hepatitis B was found to be infectious when injected i.v. into a chimpanzee up to a dilution of 10(-8). Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) were detectable in the same serum sample by radioimmunoassay up to a dilution of 10(-5) and of 10(-3), respectively. Using DNA: DNA hybridization on nitrocellulose membranes, HBV DNA sequences were detectable up to 10(-8) dilution corresponding to the infectivity level. Based on this finding, it appears that DNA: DNA hybridization is the most sensitive method for detecting hepatitis B virus (HBV) infection. In situations with low virus levels it may be the only indicator of the presence of infectious hepatitis B virus. The use of a tritium-labelled probe makes the method economical and adaptable to hospital laboratories.

摘要

在一种产生乙肝表面抗原(HBsAg)的肝癌细胞系以及急性乙型肝炎患者体内检测到了乙肝病毒(HBV)DNA。一名急性乙型肝炎患者的血清经静脉注射到黑猩猩体内,稀释至10⁻⁸时仍具有传染性。通过放射免疫测定法,在同一血清样本中分别稀释至10⁻⁵和10⁻³时仍可检测到乙肝表面抗原(HBsAg)和乙肝e抗原(HBeAg)。使用硝酸纤维素膜上的DNA:DNA杂交技术,在与感染性水平相对应的10⁻⁸稀释度下仍可检测到HBV DNA序列。基于这一发现,DNA:DNA杂交似乎是检测乙肝病毒(HBV)感染最敏感的方法。在病毒水平较低的情况下,它可能是传染性乙肝病毒存在的唯一指标。使用氚标记探针使该方法经济且适用于医院实验室。

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