Primer pair combination, detection method and use for single-plex PCR detection of aflatoxin-producing fungi

Abstract

The invention relates to the field of biotechnology and provides a single PCR detection primer pair group aiming to a fungus producing aflatoxin, a detection method, and an application thereof in detection on the fungus producing aflatoxin. In the invention, the single PCR detection primer pair group is composed of primer pairs that aim to a fungus universal molecular marker sequence (internal transcribed spacer, ITS) and three key genes, comprising an aflatoxin production regulating gene (aflR), a sterigmatocystin-o-methoxy transferase gene (omt-1), and versicolorin A dehydrogenase gene (ver-1), in the biochemically synthetic process of the aflatoxin. Through the primer pair group, technical effects of wide coverage range, strong specificity and high sensitivity during the detection of the fungus producing aflatoxin are achieved. The primer pair group is short in detection time, is simple in operation and is low in device requirement, can save large amount of manpower, materials and costs, is suitable for quick detection and is easy to promote in practical production.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a primer pair combination, detection method and application for rapid detection and identification of toxin-producing fungi by using single-plex PCR technology. Specifically, the present invention relates to primers for single-plex PCR detection of aflatoxin-producing fungi specifically targeting the fungal universal molecular identification sequence and the three key genes aflR, omt-1 and ver-1 in the biochemical synthesis process of aflatoxin For combinations, detection methods and uses. Background technique [0002] Aflatoxin (AFT) is a class of secondary metabolites with similar chemical structures mainly produced by fungi such as A.flavus, A.parasiticus and A.nomius. As many as 18 different aflatoxins have been isolated, including AFB1, AFB2, AFG1, AFG2, AFM1, AFM2, AFB2a, AFG2a, AFBM2a and AFGM2a, among which AFB1 is the most toxic and the most stable in chemical properti...

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to overcome the problems of long cycle, poor specificity and low detection sensitivity of biological detection methods for aflatoxin-producing fungi in the prior art. The present invention compares and analyzes the PCR detection method standard "SN / T 2582-2010 PCR detection method for aflatoxin-producing fungi" was optimized and designed, and primer pairs with wide coverage, strong specificity, and high sensitivity were screened out, and a single-plex PCR detection system was established by optimizing the combination, and the detection coverage was established. A detection method with a wide range, strong specificity and high sensitivity can realize efficient and accurate detection of aflatoxin-producing fungi, thereby completing the present invention

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