HRP-HBVDNA探针在临检应用中的研究

本文介绍了一种简便的检测血清ＨＢＶＤＮＡ的方法。参照Ｒｅｎｚ等人的标记方法，构建了直接酶标ＨＲＰ ＨＢＶＤＮＡ探针。此探针经与固定在硝酸纤维素滤膜上的血清靶ＤＮＡ杂交后，可通过化学发光自显影检测技术观察结果。敏感度可检测０－１ｐｇ靶ＤＮＡ，相当于同位素探针的灵敏度。对６３份ＨＢｓＡｇＨＢｅＡｇ和Ａｎｔｉ ＨＢｃＥＬＩＳＡ阳性血清以及２４份ＨＢｓＡｇＡｎｔｉ ＨＢｃ阳性，ＨｂｅＡｇ阴性血清用ＨＲＰ ＨＢＶＤＮＡ探针进行检测，结果探针ＨＢＶＤＮＡ阳性率分别为１００％（６３）和５８％（１４）；对５０份ＨＢｓＡｇ，ＥＬＩＳＡ阴性和ＡＬＴ正常的血清，探针ＨＢＶＤＮＡ全部阴性。实验结果表明本方法具有很大的推广应用价值。

Application of HRP-HBVDNA Probe in Clinic Detection

A simple method for detection of HBV DNA in sera is described in this thesis.A direct enzyme labeled HRP\|HBV DNA probe was constructed according to Renz et al.After hybridization of the probe with target DNA in sera immobilized on nitrocellulose membrane, the result could be observed by the method of chemiluminescence auto\|photographic detection.The sensitivity is as low as 0.1pg of target DNA,comparable with that of isotope probe,HBV DNA was detected in 63 sera of HBsAg,HBeAg and Anti\|HBc ELISA positive and 24 of HBsAg,Anti\|HBc positive,HBeAg negative with HBV DNA positive rates of 100% (63) and 58% (14),respectively;50 sera of HBsAg ELISA negative and ALT normal with HBV DNA all negative by the probe.The result shows that this method is of great value to popularize.