固相杂交检测HCV RNA PCR产物方法的建立

目的建立简便、快速的丙型肝炎病毒基因扩增产物的固相杂交检测方法．方法将标记有生物素的寡核苷酸引物所扩增的丙肝病毒基因产物，与通过结合在微孔反应板上的特异性探针进行快速杂交，然后通过辣根过氧化物酶标记的抗生物素进行酶联显色，读取光密度值．结果应用本方法对血清中丙型肝炎病毒核酸检测，灵敏度可达５拷贝～５０拷贝／反应．结论此方法简便、快速、特异性好、敏感性高、结果判定指标客观，可广泛应用于病毒感染的临床诊断和疗效评价及其他基因的扩增检测和基因分型等

Solid phase hybridization detection of HCV RNA PCR products

AIM To establish a simple, rapid polymerase chain reaction (PCR) based DNA amplification technique for detecting HCV RNA PCR products. METHODS For detecting HCV RNA, the primer labeled with biotin was used to amplify viral gene fragment. The 1st PCR product complemently hybridized with the specific probe covalently coupled onto microplate wells. StreptavidinePOD was used in colorimetric detection. RESULTS The sensitivity of the detection system is 5 copies50 copies of hepatitis C genome respectively. CONCLUSION The method is simple and rapid with high specificity, sensitivity and semiquantification. It can be generally applied in detection of foreign pathogen in blood, body fluid and tissue samples, in clinical diagnosis and evaluation of antiviral therapeutic effect.