Novel kit for rapidly detecting bacterial mcr-1 gene and detection method of kit

Abstract

The invention discloses a novel kit for rapidly detecting bacterial mcr-1 gene and a detection method of the kit. The kit contains reaction tubes of a loop-mediated isothermal amplification reaction liquid, wherein each reaction tube contains 20 mM of Tris-HCl with the pH being 8.8, 0.1% Tween 20, 2 mM of MgSO4, 10 mM of (NH4)2SO4, 50 mM of KCl, 1 mM of dNTP, 1.2 M of betaine, 8U of Bst DNA largepolymerase fragments, 1.6 mu M of an inner primer FIP, 1.6 mu M of an inner primer, 0.4 mu M of an outer primer F3 and an outer primer B3, 0.4 mu M of a loop primer LB and sterile double distilled water. The method of the kit comprises steps as follows: S1, bacterial genome DNA of a to-be-detected sample is extracted; S2, detection is performed with the kit; S3, if the color is green, a bacterialpolymyxin drug-resistant gene mcr-1 is positive, and if the color is orange, the bacterial polymyxin drug-resistant gene mcr-1 is negative. The kit and the method have the benefits as follows: the kithas the advantages of being high in sensitivity, high in specificity, convenient to implement, low in cost and capable of realizing high-throughput detection and the like, and the kit can be appliedto rapid detection of mcr-1 gene drug-resistant bacteria in the veterinary and human medicine fields including laboratories, farms and small animal hospital scenes.

Description

technical field [0001] The invention relates to a kit for detecting genes and a detection method thereof, in particular to a new kit for rapidly detecting bacterial mcr-1 gene and a detection method thereof. Background technique [0002] In recent years, due to the large or even inappropriate use of antibiotics in clinical practice, under the selection pressure of antibiotics, drug-resistant strains of clinical disease-related bacteria have emerged continuously, and multi-drug-resistant strains have emerged. Since drug-resistant bacteria can spread at a high rate through multiple channels, In particular, the cross-infection of humans and animals has seriously threatened the safety of human life, and at the same time greatly increased the difficulty of clinical anti-infection. In recent years, enterobacteriaceae bacteria resistant to carbapenem antibiotics have gradually appeared to aggravate this threat. Carbapenem antibiotics (imipenem, meropenem, etc.) are effective in the...

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a new rapid detection kit for bacterial mcr-1 gene and its detection in order to solve the problems of high false positive rate, high detection cost, complicated operation and long detection time in current drug resistance gene detection method

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