Molecular motor biosensor kit for detecting hepatitis A virus

Abstract

The invention relates to a molecular motor biosensor kit for detecting hepatitis A virus, belonging to the field of quick detection of food-borne viruses. The invention mainly solves the problems of long culture and multiplication cycle, low multiplication capacity, no formation of cytopathic effect, and difficulty in satisfying the detection requirements in the traditional detection method. The core technique is to utilize an F0F1-ATPase molecular motor biosensor on chromatophore; and since the F0F1-ATPase can quickly rotate, coupling between the catalytic site and the proton transfer is established through the rotation. A common sequence specific probe for various types of hepatitis A virus is connected to an epsilon subunit of the ATPase; after a sample to be detected and a negative control are respectively combined with the biosensor, the ATP synthesis amounts under the catalytic action after 10 minutes are compared; and the ATP synthesis amount can be measured according to the amount of H<+> in the environment, and the amount of H<+> is acquired according to the fluorescence intensity embodied by the H-DHPE. The method has the characteristics of short reaction time, sufficient combination with antigen antibody and the like, is convenient to operate and use, shortens the reaction time and enhances the detection sensitivity. The kit can be used for quickly and sensitively detecting hepatitis A virus in a food sample to be detected at high flux.

Description

technical field [0001] The invention relates to a rapid detection technology for food-borne viruses. Background technique [0002] The traditional detection method of hepatitis A virus (HAV) is through cell culture, but HAV not only proliferates for a long time, has low proliferative ability, but also does not form cytopathic changes, so it is difficult to meet the detection requirements. Detection techniques such as RT-PCR and ICC / PCR (integrated cell culture, PCR) established in the 1980s have improved the sensitivity and specificity of detection, and have a good application in the monitoring of hepatitis A pathogens, but the detection time still needs 5 -6h, and it is easy to be contaminated by PCR amplification products, which may cause false positives. This type of technology still needs to judge the detection results based on radioactive substance-labeled RNA probes or specific RNA bands in gel electrophoresis, and the whole process Steps such as amplification and ele...

AI Technical Summary

Problems solved by technology

[0002] The traditional detection method of hepatitis A virus (HAV) is through cell culture, but hepatitis A virus not only proliferates for a long time, has low proliferative ability, but also does not form cytopathic changes, so it is difficult to meet the detection requirements

Detection techniques such as RT-PCR and ICC / PCR (integrated cell culture, PCR) established in the 1980s have improved the sensitivity and specificity of detection, and have a good application in the monitoring of hepatitis A pathogens, but the detection time still needs 5 -6h, and it is easy to be contaminated by PCR amplification products, which may cause false positives. This type of technology still needs to judge the detection results based on radioactive substance-labeled RNA probes or specific RNA bands in gel electrophoresis, and the whole process Amplification, electrophoresis and other steps are required, and the operation is cumbersome

Not only time-consuming and labor-intensive, but also brings great inconvenience to use

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