HRP-HBVDNA探针在临检应用中的研究

Application of HRP-HBVDNA Probe in Clinic Detection

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ＨＢｓＡｇ ＨＢｅＡｇ 和ＡｎｔｉＨＢｃ ＥＬＩＳＡ 阳性血清以及２４ 份ＨＢｓＡｇ ＡｎｔｉＨＢｃ 阳性，ＨｂｅＡｇ 阴性血清用ＨＲＰＨＢＶ ＤＮＡ 探针进行检测，结果探针ＨＢＶ ＤＮＡ 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ＨＢＶ ＤＮＡ 全部阴性。实验结果表明本方法具有很大的推广应用价值。

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.\;