Method for preparing detection chip and method for detecting pathogens using the chip

Abstract

The invention discloses a method for preparing a detection chip, which comprises the following steps: 1) designing a bacterial oligonucleotide probe; 2) preparing the chip. The present invention also discloses a method for detecting pathogens in a plurality of food samples by using the detection chip at the same time, comprising the following steps: 1) designing primers; 2) extracting DNA from samples to be tested to obtain extracts; 3) multiplex PCR of target genes Amplification; 4) chip hybridization detection; 5) detection and analysis of hybridization results. The detection method of the invention has simple steps, is easy to operate, and has a high detection result accuracy rate.

Description

Technical field [0001] The invention belongs to the detection technology of microorganisms, especially the nucleic acid detection technology of pathogenic microorganisms in milk and dairy products; in particular, it relates to a method and chip used for detecting pathogens in 12 food samples at the same time. Background technique [0002] Food-borne pathogens often cause mass injuries and cause certain harm to people's health, lives and property. According to an incomplete statistical report of the World Health Organization (WHO), 2.1 million people worldwide died of diarrhea in 2000, mostly due to the contamination of pathogenic bacteria in food and drinking water. According to statistics from the Ministry of Health, a total of 150 major food reports were received in 2000, resulting in 6,237 people being poisoned and 135 deaths; in 2001, a total of 185 major food poisoning incidents occurred, resulting in 15,715 people being poisoned and 146 deaths. [0003] The basis for judging...

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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