A rapid detection kit for goat infectious pleuropneumonia and its preparation method

Abstract

The invention discloses a rapid detection kit for goat contagious pleuropneumonia and a preparation method thereof. The kit is established on the basis of molecular biology. The kit comprises 2*loop-mediated isothermal amplification (LAMP) reaction liquid, a primer mixer, BstDNA polymerase, a color-developing agent, a standard positive template and ddH2O, wherein the primer mixture comprises a forward inner primer FIP, a downward inner primer BIP, a forward outer primer F3 and a downward outer primer B3; and the ratio of the FIP to the BIP to the F3 to the B3 is 8:8:1:1. The kit has the characteristics of high sensitivity and specificity and the like, can react quickly, is easy to operate, does not need special instrument equipment and overcomes the defects of long pathogeny separation time, high workload, fussy detection method, complicated operation and the like of goat contagious pleuropneumonia, and the detection method is complicated, is difficult to operate and the like, and can be used for detecting whether clinical samples are infected with Mycoplasma capricolum subsp. Capripneumonia as well as performing early diagnosis and molecular epidemiological survey of goat contagious pleuropneumonia.

Description

technical field The invention relates to a loop-mediated isothermal amplification technique for detecting the genome DNA of Mycoplasma capricum subspecies goat pneumonia, specifically a rapid detection kit for goat infectious pleuropneumonia, and the invention also includes a preparation method of the kit. Background technique The commonly used methods for diagnosing goat infectious pleuropneumonia include agglutination and PCR, and these methods have certain shortcomings. The agglutination test is mainly the indirect hemagglutination method. The disadvantage of this method is that the detection sensitivity is low, and there will be cases of missed detection; moreover, this method is aimed at the detection of goat infectious pleuropneumonia antibodies. The positive animals may be caused by vaccination with mycoplasma vaccine. of. Both PCR and LAMP methods are sensitive, specific and efficient new diagnostic techniques based on molecular biology. However, the PCR process of...

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Problems solved by technology

However, the PCR process of goat infectious pleuropneumonia is relatively complicated. After the amplification, it needs to be identified by enzyme digestion to determine whether it is positive, and this method needs an expensive PCR instrument to cooperate with it. The reaction time is long, and agarose gelation is required after the reaction. Gel electrophoresis uses a gel imager to observe the results, and in the process of animal genome DNA extraction, there may be some remaining inhibitory factors of the PCR reaction that affect its specificity and sensitivity

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