Mycoplasma hyopneumoniae loop-mediated isothermal amplification kit and application thereof

Abstract

The invention discloses a mycoplasma hyopneumoniae loop-mediated isothermal amplification kit and the application thereof. The kit comprises LAMP primers, 2x reaction buffer, Bst DNA polymerase, a fluorescence visual detection reagent, ultrapure water and a mycoplasma hyopneumoniae DNA template, wherein the LAMP primers include outer primers F3 and B3, inner primers FIP and BIP, and loop primers LF and LB. The kit can be applied to detection of lesion tissue of suspected mycoplasma pneumoniae of swine and a mycoplasma hyopneumoniae culture. Specificity detection and sensitivity detection prove that by the adoption of the LAMP detection method, reaction can be monitored in real time, the mycoplasma hyopneumoniae copy number can be detected, a detection result can be obtained quickly and accurately, and convenience is brought to easy, quick and reliable detection of mycoplasma hyopneumoniae.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a fast, visualized and real-time quantitative detection of Mycoplasma hyopneumoniae, a loop-mediated isothermal amplification kit and an application thereof. Background technique [0002] Mycoplasma hyopneumoniae (Mycoplasmahyopneumoniae, Mhp) is an important pathogen that causes mycoplasma pneumoniae (mycoplasmalpneumoniaofswine, MPS), also known as swine asthma. It was first isolated from a cell-free medium by Goodwin (1964), and then Shanghai Academy of Agricultural Sciences The Institute of Animal Husbandry and Veterinary Medicine isolated the pathogenic strain for the first time in 1973, which mainly causes chronic contact respiratory infectious diseases in pigs with cough and wheezing as the main symptoms. [0003] Mhp can be transmitted through the air, and it is difficult to isolate and block its transmission route, resulting in a high incidence of swine asthm...

AI Technical Summary

Problems solved by technology

Among the existing pathogen detection methods for Mycoplasma hyopneumoniae, pathogen isolation is the most accurate method for the diagnosis of Mycoplasma hyopneumoniae pneumonia, but at present, the isolation, culture and identification of Mycoplasma hyopneumoniae in vitro is difficult, low-sensitivity, time-consuming, and requires complex equipment and culture conditions, so in developing countries and areas with limited clinical conditions in my country, it is particularly important to establish a simple and rapid diagnostic method

Although the polymerase chain reaction (PCR) method has high specificity and sensitivity, it requires expensive PCR machines, electrophoresis tanks and other precision instruments, and is not suitable for grassroots development.

Images