Method for detecting aquatic product pathogenic microorganisms based on random amplifying and labelling and in-situ synthesized microfluid chip

Abstract

The invention discloses an in-situ synthesized microfluid chip. The in-situ synthesized microfluid chip has 895 oligonucleotide probes and contains 42 positive / negative control probes, which are as shown in a table 1. Besides, the invention further provides a method for detecting aquatic product pathogenic microorganisms through the in-situ synthesized microfluid chip. The method comprises the following steps of preparing oligonucleotide chips, labelling genetic fragments of the pathogenic microorganisms by a random amplifying method, hybridizing the chips, and according to the hybridization result, judging whether samples contain the aquatic product pathogenic microorganisms or not. The high-flux detection capacity of the chips and the simple, convenient and efficient characteristics of the random amplifying method are utilized, and quick parallel detection is performed on fragments of conservative genes and variation genes of main pathogenic microorganisms of aquaculture animals in China.

Description

technical field [0001] The invention relates to a detection and identification method for aquatic pathogenic microorganisms, in particular to a method for detecting aquatic pathogenic microorganisms by using a random amplification labeling method and in-situ synthesis microfluidic chip technology. Background technique [0002] In recent years, my country's fishery production has achieved rapid development, and its proportion in agricultural output value has risen steadily. It has become one of the four pillar industries of modern agriculture (grain, meat, aquatic products and poultry eggs). The aquaculture industry has also shown an unprecedented improvement in the scale of farming and farming technology, especially in the rapid development of intensive farming and factory farming. However, under the current high-density, intensive, and multi-species aquaculture environment and mode, various aquaculture animal diseases occur frequently, causing heavy losses to farmers, and h...

AI Technical Summary

Problems solved by technology

[0003] The frequent occurrence of aquatic diseases has led to the prominent problem of fishery drug abuse, which not only seriously affects the quality and safety of aquatic products, but also causes more serious environmental pollution problems

In addition, the mutation and recombination of pathogenic bacteria and viruses have greatly increased, as well as the emergence of new biologically derived diseases.

For example, in recent years, the relatively serious "stealing death disease" ("death disease") in the shrimp farming process has caused the most serious disaster in the 20-year history of the vannamei farming industry in South China in 2010 (the death rate reached more than 80%). Now people still don't know the cause of it, but the more popular theory is Vibrio infection, outbreak of new virus or deterioration of aquaculture water environment

This reflects that the traditional isolation and cultivation of pathogenic microorganisms and observation of symptoms are difficult to meet the actual production needs.

Moreover, the detection and diagnosis of a single (or a small number of) pathogens cannot fully reflect the cause and status of the disease, which is not conducive to formulating effective control measures.

Images