【摘要】 <正> 近三、四年来,DNA探针和分子杂交技术已开始应用于各种病毒或细菌感染、癌症和遗传病的诊断上。它具有准确性强、敏感度高的特点。但目前所用探针大都使用³²P,不可避免具有同位素的缺点,效期短、需要特殊的防护设备和污物处理。目前许多人研制效期长的非同位素的生物素探针,但尚未达到实用地步。我们实验室用缺口翻译法将生物素化核苷更多还原

【Abstract】 HBV/adw DNA clone pAM6 (kindly gifted by Dr. J. Gerin) was linearized by Sal I digestion and further purified by gel electrophoresis. We nick-translated the DNA with Biotin-11-dUTP. For dot hybridization we used a Bio-Rad Bio-Dot apparatus and NC or Zeta-probe membrane. After hybridization we used streptavidine-alkaline phosphatase, NBT and BCIP for visualization. By using our biotin probe we can detect as little as 0.66 Pg of HBV DNA. A comparison study of 147 sera from liver diseases using our probe and commercial 32P probe showed that the biotin probe pickup 90 positives while 32P probe pickup 66 positives. The lowered sensitivity of 32P probe may be due to deterioration of the kit during transportation. Analysis of 55 cases of HBsAg positive sera in relation to HBV DNA (biotin), HBeAg and anti-HBe showed that the No. of DNA positives was 41 among 50 HBsAg positive HBeAg positive cases (82%) and 2 among 5 HBsAg positive anti-HBe positive cases. In a study of 71 cases of liver diseases, the No. of DNA positive was 12 among 25 primary liver carcinoma (48%). 16 among 26 chronic carriers (61.5%),3 among 3 hepatitis gravis, 2 among 8 chronic liver diseases including cirrhosis, 0 among 7 extrahepatic cancer or metastatic liver cancer. There was no HBV DNA positive in 2 cases of non-hepatic disease and in 23 normal persons.更多还原