一种斑点杂交检测方法

Dot hybridization detection method

Abstract

本发明适用于病毒检测领域,提供了一种斑点杂交检测方法,所述方法包括:将含有DNA样品的杂交膜封入杂交袋中并加入杂交溶液,封闭杂交膜上多余的非特异性DNA结合位点;将所述杂交膜置于烧杯,加入冲洗溶液于摇床中洗膜,洗去未结合的探针并封闭所述杂交膜上非特异性的蛋白质结合位点;取出所述杂交膜置于平皿中,加入含有抗地高辛抗体的马来酸缓冲液中,使酶联抗体与地高辛相结合并洗去未结合的酶联抗体;将所述杂交膜置于显色液中显色,使用无酶水终止反应。本发明可同时检测多个样品,操作方便,节省时间,无需特殊仪器设备,阳性信号清晰,试剂成本较低,可做半定量分析并可检测出病毒载量低的病毒。
The invention is applicable to the field of virus detection and provides a dot hybridization detection method. The method comprises the following steps of: packaging a hybridized film containing a DNA (Deoxyribonucleic Acid) sample into a hybridization bag and adding a hybridization solution; closing surplus non-specific DNA combined sites on the closed hybridized film; putting the hybridized film into a beaker and adding a washing solution to wash the film in a shaking table to wash off an uncombined probe and close the non-specific protein combined sites on the hybridized film; taking out the hybridized film and putting the hybridized film into a flat vessel; adding a maleic acid buffering solution containing an anti-digoxin antibody to combine an enzyme coupled antibody and wash off an uncombined enzyme coupled antibody; and placing the hybridized film into a color developing solution to develop the color and utilizing non-enzyme water to stop the reaction. According to the dot hybridization detection method, various samples can be detected at the same time, the operation is convenient and the time is saved; special instruments and equipment are not needed, a positive signal is clear and the reagent cost is low; and a semi-quantitative analysis can be carried out and viruses with a low virus load can be detected.

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Patent Citations (3)

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    Title
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