Preparation of Alkaline Phosphatase Labeled HBV Probe and Its Application in Serum Detection

TU Zhi-Meng; KE Li-Hua

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April 1991

TU Zhi-Meng and KE Li-Hua. Preparation of Alkaline Phosphatase Labeled HBV Probe and Its Application in Serum Detection[J]. Virologica Sinica, 1991, 6(2).

Citation: TU Zhi-Meng, KE Li-Hua. Preparation of Alkaline Phosphatase Labeled HBV Probe and Its Application in Serum Detection .VIROLOGICA SINICA, 1991, 6(2) : 132.

酶标HBV DNA探针的研制与应用

涂知明 ,
柯丽华

1.
中国科学院武汉病毒研究所
2.
中国科学院武汉病毒研究所武汉 430071湖北医学院生物教研室

摘要

本实验采用一种非放射性物质——碱性磷酸酶标记乙肝病毒HBV DNA制备分子探针。碱性磷酸酶在苯醌作用下与单链DNA联结,形成DNA和酶的共价复合物,即酶标探针。此探针通过分子杂交与待测DNA结合,与酶的底物作用显色,几小时内可观察结果,其最低检测量约为10pg。用此探针检测乙肝病人血清中的HBV DNA,与~(32)P标记的探针比较,酶标探针可检测出~(32)P标记探针检出率的95.7％。结果表明,所合成的酶标探针具有准确、简便、快速、安全而经济的优点,具有应用前景。
- 碱性磷酸酶
- , 酶标探针
- , 乙肝病毒DNA

Preparation of Alkaline Phosphatase Labeled HBV Probe and Its Application in Serum Detection

TU Zhi-Meng ,
KE Li-Hua

1.
Wuhan Institute of Virology
2.
Academia Sinica

Abstract

A method using nonradioactive material—alkaline phosphatase to label HBV DNA as probe has been studied and used in clinical experiments to detect the HBV DNA in hepatitis patient s serum. Alkaline phosphatase coupled with PEI (polyethyleneimine) using P-benzoquine as cross-linking reagent. The modified phosphatase was covalently linked to single stranded DNA using glutral dehyde. Such single stranded DNA enzyme complexes have been tested for blot hybridization, after hybridization and incubation with a subs...
- Alkaline phosphatase Probe Cross-linking Hepatitis

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Preparation of Alkaline Phosphatase Labeled HBV Probe and Its Application in Serum Detection

TU Zhi-Meng ,
KE Li-Hua

1. Wuhan Institute of Virology
2. Academia Sinica

Abstract: A method using nonradioactive material—alkaline phosphatase to label HBV DNA as probe has been studied and used in clinical experiments to detect the HBV DNA in hepatitis patient s serum. Alkaline phosphatase coupled with PEI (polyethyleneimine) using P-benzoquine as cross-linking reagent. The modified phosphatase was covalently linked to single stranded DNA using glutral dehyde. Such single stranded DNA enzyme complexes have been tested for blot hybridization, after hybridization and incubation with a subs...