PCR detection kit and detection method for Streptococcus equi

Abstract

The present invention relates to a PCR detection kit for detecting Streptococcus equi (further called Streptococcus equi subsp equi). The detection kit comprises primers, dNTP, a buffer and DNA polymerase, wherein the upstream primer is 5'-CTATTAAAGTCTCCATTG-3', and the downstream primer is 5'-AATGTTGTTCAAGCAAATTC-3'. The PCR detection method comprises: providing a sample template requiring detection; adding dNTP, a 10*buffer, primers, DNA polymerase, a sample requiring detection and ddH2O to a PCR thin-walled tube, and uniformly mixing; amplifying the mixture in the thin-walled tube through a PCR instrument; carrying out electrophoresis on the amplification product in electrophoresis equipment; and analyzing and judging the result. According to the present invention, the PCR kit preparation method is simple and is suitable for industrial production; the PCR detection method has characteristics of high Streptococcus equi detection sensitivity, short detection period, rapidness, and strong operability, wherein detection accuracy can achieve template DNA of 80 bacterial while a detection cost is relatively low; and the detection kit and the detection method have important application values in clinical diagnosis of the disease.

Description

technical field [0001] The invention belongs to the field of microbial biotechnology, and the specific content relates to a PCR detection kit which can be used to detect Streptococcus equi (also known as Streptococcus equi subspecies equi), comprising primers, dNTP, buffer and DNA polymerase, The upstream primer is 5'-CTATTAAAGTCTCCATTG-3', and the downstream primer is 5'-AATGTTGTTCAAGCAAAATTC-3'; PCR detection method: provide the sample template to be tested; add dNTP, 10×buffer, primers, DNA to the PCR thin wall tube Polymerase, test sample and ddH 2 O, mix well; make the mixture in the thin-walled tube amplify on the PCR instrument; electrophoresis the amplified product in the electrophoresis device; analyze and judge the result. technical background [0002] Equine Streptococcus (Streptococcus. Suppurative lymphangitis of the head. Therefore, research on S. equine is urgently needed. The United States, Britain and other countries have always attached great importance...

AI Technical Summary

Problems solved by technology

[0006] At present, there are very few domestic studies on the diagnosis of equine gland disease. The traditional methods of isolation and culture, biochemical identification, and serotyping are still widely used in the detection of this bacteria, which is time-consuming and labor-intensive, and is not suitable for the detection needs in production.

There are also studies that use 16SrDNA technology as an auxiliary diagnosis. This diagnostic technology still needs to be verified by sequencing and sequence analysis, and the specificity is not strong, which prolongs the detection cycle (Chen Xinhua, Kang Lichao, Huang Xin, etc., Isolation and Identification of Streptococcus equine epidemic , Chinese Journal of Veterinary Medicine, 2013 1(49): 38-39), which cannot meet the needs of rapid, simple and accurate diagnostic methods for the disease in current clinical and production practices

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