Kit for loop-mediated isothermal amplification detection of Mycoplasma ovipneumoniae and preparation and usage methods thereof

Abstract

The invention discloses a raw material composition of a kit for loop-mediated isothermal amplification (LAMP) detection of Mycoplasma ovipneumoniae and a preparation method and a usage method of the kit. The raw material composition comprises 1000-2000muL of reaction solution, 100-200muL of Bst (Bacillus stearothermophilus) DNA polymerase, 50-100muL of positive control, 50-100muL of negative control, 1-2mL of liquid paraffin and 1-2mL of ultrapure water. The reaction solution comprises an inner primer mixture solution, an outer primer mixture solution, an LAMP reaction buffer, Mg<2+> and dNTPs (deoxyribonucleotide triphosphates), wherein the volume ratio of the five liquids in the reaction solution is 2:2:2.5:2:1. The preparation method comprises the following steps: 1) determination of an optimum reaction temperature and an optimum reaction time; 2) specific detection, sensibility test and clinical application detection and 3) kit packaging. The kit disclosed by the invention has rapid, simple and accurate characteristics for pathogen detection of goat suspected cases of mycoplasma ovipneumoniae infection in goat farms.

Description

technical field [0001] The invention relates to a Mycoplasma pneumoniae ring-mediated isothermal amplification detection kit based on Mycoplasma ovipneumoniae 16S rRNA gene and a preparation and use method. Background technique [0002] Mycoplasma ovis pneumoniae ( Mycoplasma ovipneumoniae , Mo) for goat infectious pleuropneumonia ( Contagious Caprine Plneuropneumonia, CCPP) is one of the main pathogens, which can infect sheep and goats. After the infection of goats, the clinical manifestations are respiratory disorder, emaciation, high fever, lying down and other symptoms, and the mortality rate is high, causing serious economic losses to the goat breeding industry. [0003] At present, the detection of Mo at home and abroad mainly relies on the isolation and identification of pathogens, serological tests and polymerase chain reaction techniques. Compared with the three diagnostic methods, the traditional separation and identification of mycoplasma has high requirements ...

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a ring-mediated isothermal amplification detection kit for Mycoplasma ovis pneumoniae and its preparation and use methods, which can avoid the blindness and cumbersomeness of clinical pathogen detection for suspected cases of Mycoplasma ovis pneumoniae infection in goats, And reduce the high cost and high equipment requirements for the detection of Mycoplasma pneumoniae by polymerase chain reaction technology, and provide technical support for the prevention and treatment of Mycoplasma pneumoniae from goats, so as to overcome the high cost, difficulty and time-consuming diagnosis of existing technologies. many deficiencies

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