LAMP primer combination for detecting 8 environmental pathogens of dairy cow mastitis and application thereof

Abstract

The invention discloses an LAMP primer combination for detecting 8 environmental pathogens of dairy cow mastitis and application thereof. The primer combination provided by the invention is composed of 48 primers shown in sequence 1 to sequence 48. The primer combination provided by the invention can be used for detecting whether a to-be-detected bacterium is Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus dysgalactiae, Streptococcus uberis, Salmonella typhimurium, Proteus mirabilis or Candida albicans, and can be used for detecting whether a to-be-detected sample contains Escherichia coli and / or Klebsiella pneumonia and / or Pseudomonas aeruginosa and / or Streptococcus dysgalactiae and / or Streptococcus uberis and / or Salmonella typhimurium and / or Proteus mirabilis and / or Candida albicans. The primer combination provided by the invention is used for identifying and detecting the 8 environmental pathogens of dairy cow mastitis, has high specificity and high sensitivity, and can realize simple, rapid and accurate detection, thus having great popularization value.

Description

technical field [0001] The invention relates to a combination of LAMP primers for detecting eight environmental pathogens of cow mastitis and its application. Background technique [0002] Mastitis in dairy cows is inflammation of the lactating tissue in the milk area, usually caused by bacterial infection, and occurs in one or more milk areas, resulting in reduced milk production, cessation of milk production, and changes in milk quality. Dairy mastitis affects 25-50% of dairy cows every year and is the most costly disease in dairy farming. [0003] Cow mastitis is caused by a variety of non-specific pathogenic microorganisms. At present, there are about 150 kinds (including cocci, bacilli, mycoplasma, fungi or yeasts, viruses, etc.), among which the more common ones can be divided into infectious pathogens and non-communicable pathogens. According to reports, the annual loss caused by cow mastitis in the world is 2 billion US dollars in the United States, 267 million in ...

AI Technical Summary

Problems solved by technology

Traditional methods for detecting pathogenic bacteria such as isolation culture, morphological observation, physiology and biochemistry, and selective medium have the characteristics of low sensitivity, poor specificity, heavy workload, long time-consuming, and low throughput.

The microbial culture method takes a long time and needs 3-4 days to produce a test report, which cannot meet the purpose of timely treating bovine mastitis against pathogens

Immunological techniques (such as ELISA) have high sensitivity, but are prone to contamination and often have false positives

Usually, a specific primer for one pathogenic bacteria and one specific primer is used to check the specific pathogen. Although it can achieve the purpose of rapid, correct and convenient diagnosis of a single bacterium, when the pathogen is not named, it needs a variety of different primers for detection. test, which is clearly not ideal for the complexity of pathogens and the diversity of PCR programs

PCR has the disadvantages of long detection time, easy contamination, and high false positive rate, which limits its application.

There are many technical problems in the above technologies and methods for the detection of pathogenic microorganisms, and in practice, due to the wide variety of pathogenic microorganisms, it is difficult for the above methods to efficiently detect multiple pathogenic microorganisms at the same time

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