Treble PCR method for simultaneously detecting mycoplasma hyopneumoniae, porcine pasteurella multocida and haemophilus parasuis

Abstract

The invention discloses a treble PCR method for simultaneously detecting mycoplasma hyopneumoniae, porcine pasteurella multocida and haemophilus parasuis. A treble PCR detection method for directly detecting 3 pathogens once from a sample is established through the following steps: firstly, screening out conservative genetic fragments with the gene type characteristics of the pathogens, using the conservative genetic fragments as 3 gene target points for PCR detection, respectively synthetizing and amplifying primers of the target point genes, and then putting 3 pairs of the primers of the 3 genetic fragments in a PCR reaction system. Through the adoption of the treble PCR method disclosed by the invention, on one hand, the pathogens can be accurately detected, and a mixed infection condition can be analyzed, so that the epidemic and development trend of an epidemic situation can be controlled, and the treble PCR method has double effects; on the other hand, compared with the conventional PCR method, the detecting time is shortened by 24 hours, so that the purpose of quickly detecting actual samples is achieved, and the cost is reduced.

Description

Technical field [0001] The invention relates to a multiple PCR detection method in the field of biotechnology, in particular to a triple PCR method capable of simultaneously detecting Mycoplasma hyopneumoniae, Pasteurella multocida and Haemophilus parasuis. Background technique [0002] Porcine respiratory diseases have become one of the important mass diseases affecting the pig industry. Studies have shown that swine respiratory diseases are diseases caused by mixed infections of one or more bacteria and viruses. Usually viruses or mycoplasma invade the respiratory tract first, destroying the defense barrier, and various airborne pathogens such as Pasteurella multocida and Haemophilus parasuis enter the respiratory tract and lungs to cause secondary or mixed infections. Mycoplasma hyopneumoniae (Mhp) is one of the main primary pathogens. As pigs are infected with MPS, it is difficult to cure and the disease is difficult to purify. Avoiding contact with MPS is the most effective...

AI Technical Summary

Problems solved by technology

Traditional methods such as morphological observation, physiological and biochemical identification, and immunological identification are mostly used for the identification of Pasteurella. These methods are time-consuming and labor-intensive and have poor accuracy. Therefore, it is difficult to judge the pathogen only by conventional inspection methods and clinical experience. , which will delay the best time for the prevention and control of animal diseases; Haemophilus parasuis is a Gram-negative bacterium that can cause fibrous serositis, meningitis and arthritis in pigs, and is often associated with pigs related to respiratory disease

Haemophilus parasuis is one of the pathogenic bacteria that is more difficult to cultivate than Mycoplasma hyopneumoniae. The current isolation and detection of it is more than 20% less than the actual infection rate of the farm. It is usually associated with Mycoplasma hyopneumoniae and Pasteurella multocida. , pig reproductive and respiratory viruses and other pathogenic bacteria mixed infection, due to the harsh culture conditions, the research on it is limited

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