Method for preparing detection chip and method for detecting pathogen by using the same

Abstract

The invention discloses a preparation of a detection chip, comprising the steps of: 1) designing oligonucleotide probe of bacteria; 2) preparing the chip. The invention further discloses a method for detecting pathogene in multiple food samples with the detection chip, comprising the following steps: 1) designing a primer; 2) extracting DNA from samples to be detected, and getting extracting solution; 3) multi-PCR amplifying of target gene; 4) hybridization detecting of chip; and 5) detecting and analyzing of hybridization result. The detection step of the invention is simple, easy to operate, and produces high correct rate result.

Description

technical field [0001] The invention belongs to microorganism detection technology, in particular to the nucleic acid detection technology of pathogenic microorganisms in milk and dairy products; in particular, it relates to a method capable of simultaneously detecting pathogens in 12 food samples and a chip used therefor. Background technique [0002] Foodborne pathogenic bacteria often cause mass damage, causing certain harm to people's health, life and property. According to the incomplete statistical report of the World Health Organization (WHO), 2.1 million people died of diarrhea in the world in 2000, most of which were attributed to the contamination of pathogenic bacteria in food and drinking water. According to statistics from the Ministry of Health, in 2000 there were 150 major food poisoning reports, resulting in 6237 poisonings and 135 deaths; in 2001 there were 185 major food poisoning incidents, resulting in 15715 poisonings and 146 deaths. [0003] The basis ...

AI Technical Summary

Problems solved by technology

At present, there are already some chips used for food testing. However, these chips also have the following shortcomings and deficiencies: 1. Although high-throughput detection of pathogens can be achieved, generally one chip can only detect all pathogens in one sample; 2. At present, there are generally two types of fluorescent labels. One is to add Cy-dCTP while amplifying. However, since Cy is a large molecule and has a large steric hindrance, it is not easy to combine during the chain extension process; the other The first method is to synthesize primers with fluorescent groups first. This method also has relatively large disadvantages: first, primers with fluorescence are not easy to store, and fluorescence quenching will occur after a long time; secondly, the price of synthesizing primers with fluorescence is relatively low. Expensive, and different primers are needed to amplify different genes, so many fluorescent primers need to be synthesized; finally, because the primers are fluorescent, once purified, or the washing steps are not done well, false positives will appear in the results, or Locally high background signal, etc.

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