应用地高辛标记HCMV DNA混合探针检测人巨细胞病毒DNA  

Detection of Human Cytomegalovirus DNA Using Mixed Probes Labelled with Digoxigenin

作　　者：王玲[1] 肖卫华[1] 董伟[1] 程兆军[1] 胡嘉第 徐德启[1] 

机构地区：[1]卫生部长春生物制品研究所,长春130062

出　　处：《中国生物制品学杂志》1992年第4期164-166,共3页Chinese Journal of Biologicals

摘　　要：本文比较了^(32)P和地高辛标记的HCMV EcoRI单一Z片段与EcoR IZ、K、J混合片段探针的敏感性。结果^(32)P和地高辛分别标记的EcoRIZ单一片段检测敏感性分别为50pg和1pg,而两者标记的EcoRI Z、K、J混合片段检测敏感性分别提高到5pg和0.1pg。地高辛标记的混合片段探针比^(32)P标记单一EcoRIZ片段探针敏感性提高500倍,比^(32)P标记混合探针高50倍,比地高辛标记的EcoRI Z片段探针高10倍。应用地高辛标记混合片段探针检测了正常小儿尿标本10份,阳性率为60％(6/10),与免疫组化方法对比,两者符合率为100％。检测肾移植患者尿标本33份,阳性率为46％(15/33);白血病患者尿标本18份,阳性率为72％(13/18),与PCR方法对比两者符合率为93％。地高辛标记混合探针可快速、敏感、特异地检测人巨细胞病毒感染。The sensitivity of probe EcoRI Z fragment and mixed probes EcoRI Z, K, J fragments of HCMV DNA labelled with digoxigenin (dig.) by random hexanucleotide priming were compared with the two probes labelled with ^(32)p by nick translation. The sensitivity of probe EcoRI Z labelled with ^(32)P and dig. were 50pg and 1pg of homologous DNA, respectively. The sensitivity of mixed probes EcoRI Z, K, I labelled with both methods were 0.1pg and 5pg of homologous DNA, respectively. The sensitivity of mixed probes labelled with dig. increased by 500-fold over that of single probe EcoRI Z fragment labelled with ^(32)P, and 50-fold over mixed probes labelled with ^(32)P, and 10-fold over probe EcoRI Z labelled with digoxigenin. The HCMV DNA of 10 urine samples form normal children, 33 from kidney transplant recipients and 18 from patients with leukaemia were detected by using mixed probes labelled with dig.. The positive rates were 60%, 46% and 72%, respectively. Samples from normal children were also detected by immunohistochemical method, and the result completely tallyed with that obtained from dig. technique. The HCMV DNA of samples from kidney transplant recipients and patients with leukaemia were detected by PCR method and 93% of the results tallyed with that obtained from dig. technique. The above-mentioned results showed that mixed probes labelled with dig. could detect HCMV DNA rapidly, sensitively, and specifically.

关 键 词：地高辛 混合探针 巨细胞病毒DNA 

分 类 号：R977[医药卫生—药品]