SIMULTANEOUS DETECTION OF HBV AND HCV GENOMIC SEQUENCES BY PCR

通过 PCR 同时检测 HBV 和 HCV 基因组序列

• 批准号: 3792566
• 负责人: S NEDJAR
• 金额: --
• 依托单位: CENTER BIOLOGICS EVALUATION RESEARCH TRANSFUSION
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3792566
• 关键词: alanine transaminase; antiviral antibody; blood donor; communicable disease diagnosis; complementary DNA; diagnosis design /evaluation; genome; hepatitis B; hepatitis B virus group; hepatitis C virus; human tissue; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; radiotracer; serology /serodiagnosis; serum; virus DNA; virus RNA

A sensitive and specific two stage polymerase chain reaction (PCR) method was developed for the simultaneous amplification and detection of specific genomic sequences of hepatitis B virus (HBV) and hepatitis C virus (HCV) in donor sera. Initially, HCV-RNA was reverse transcribed to cDNA. This cDNA, and DNA from HBV, were then co-amplified by using primer pairs derived from conserved regions of HBV and HCV nucleotide sequences. The specificity of PCR products was confirmed by liquid hybridization analysis using 32P-end labeled oligomer probes specific for HBV and HCV nucleotide sequences. Independent sets of sera, positive and negative by PCR for either HBV-DNA or HCV-RNA, were used as controls. Nine donor sera, antibody reactive for both HBV (HBsAg and anti-HBc) and HCV (anti-HCV), were tested. Our assay detected HBV and HCV-specific genomic sequences in 9/9 sera reactive for both HBV and HCV antibodies. The ability to co-amplify and detect HBV and HCV sequences, non-isotopically may be particularly useful in the clinical laboratory setting.

一种敏感、特异的两步法聚合酶链反应（PCR） 建立了一种同时扩增和检测 B型肝炎病毒（HBV）和丙型肝炎病毒的特异性基因组序列 供者血清中的丙型肝炎病毒（HCV）。 最初，HCV-RNA被逆转录， cDNA。 然后使用PCR共扩增该cDNA和来自HBV的DNA 来自HBV和HCV核苷酸保守区的引物对 序列的 PCR产物的特异性经液相色谱法证实 使用32 P-末端标记寡聚体探针特异性杂交分析 HBV和HCV核苷酸序列。 独立的血清组， PCR检测HBV-DNA或HCV-RNA阳性和阴性，用作 对照 9份供体血清，对HBV（HBsAg和 抗-HBc）和HCV（抗-HCV）。 我们的检测方法检测到HBV， 在9/9份对HBV和HCV均反应的血清中检测到HCV特异性基因组序列 抗体的 共扩增和检测HBV和HCV序列的能力， 在临床实验室中可能特别有用 设置.