PCR detection kit for regulator gene producing aflatoxin and detection method thereof

Abstract

The invention provides a PCR detection kit for regulator gene producing aflatoxin and a detection method thereof. The detection kit mainly comprises special amplification primers, PCR buffer liquid, deoxyl nucleoside triphosphate (dNTP) mixture and DNA polymerase; the special primers is with upstream primer sequence of 5'-CGAAAGCTCCGGGATAGCTGTACG-3', downstream primer sequence of 5'-CGTCAGACAGCCACTGGACACGG-3'. The invention has the advantages: (1) early warning: the method is a rapid and accurate microbe detection technology for the food threatened by aflatoxin, an early warning way to essentially solve the problem; (2) convenient and saving time and labour: compared with the traditional toxigenic culture technology, the invention can save time and labour, and is capable of finishing detection within three days; (3) high sensitivity and strong specificity: as the toxigenic culture technology can be heavily influenced by environment, and be easy to appear false positive, the specific gene amplification technology is adopted to ensure high diagnosis sensitivity and stronger sensitivity.

Description

(1) Technical field [0001] The invention relates to a PCR detection kit for aflatoxin-producing regulatory gene nucleic acid and a detection method thereof. (2) Background technology [0002] Aflatoxin is the strongest carcinogen known so far. Its carcinogenicity is 900 times that of carcinogen dimethyl cream yellow, 75 times greater than dimethylnitrosamine inducing liver cancer, and 3,4-benzo Pyrene is 4000 times more carcinogenic. The toxicity of aflatoxin to humans, livestock, and poultry is related to its inhibition of the transmission of genetic information in cells and interference with protein synthesis. Aflatoxins also have additive effects with other human pathogenic factors (such as hepatitis virus, etc.). Between 2004 and 2005, Kenya suffered the largest outbreak of aflatoxin poisoning in history, resulting in 150 deaths. In July 2005, the U.S. Centers for Disease Control and Prevention and the World Health Organization jointly held an international seminar on...

AI Technical Summary

Problems solved by technology

[0007] At present, the PCR method has been widely used in the detection of various bacteria, but the detection of toxin-producing fungi is relatively rare. The primers for the detection of aflatoxin-producing regulatory gene aflR reported in 1996 were used in many reports, but after many The second experiment found that the amplification efficiency was unstable, and no amplification product could appear in positive samples. Further analysis found that the upstream primers of the aflatoxin-producing regulatory gene: a. easy to form primer dimers, b. analyzed by Blast of Genbank (to December 2008) Polymorphic sites appeared in 25 of 87 aflR gene records

Obviously, this pair of primers cannot meet the current needs of PCR detection of aflatoxin-producing regulatory genes

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