Primer probe combination and kit for detecting aflatoxin-producing fungi in food

A technology of aflatoxin, primer probe, applied in the field of microbial detection, can solve the problems of difficult application, long inspection cycle, cumbersome steps, etc., and achieve the effects of fast detection speed, strong detection specificity and high sensitivity

Pending Publication Date: 2022-01-14
GONGBEI CUSTOMS TECH CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both methods have the limitations of cumbersome steps and long inspection cycle, which are difficult to apply in emergency monitoring

Method used

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  • Primer probe combination and kit for detecting aflatoxin-producing fungi in food
  • Primer probe combination and kit for detecting aflatoxin-producing fungi in food
  • Primer probe combination and kit for detecting aflatoxin-producing fungi in food

Examples

Experimental program
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Effect test

Embodiment 1

[0047] A combination of primers and probes for detecting aflatoxin-producing fungi in food, said aflatoxin-producing fungi include Aspergillus flavus and Aspergillus parasitica, said primers include upstream primers and downstream primers, and the nucleotide sequence of said upstream primers is as SEQID As shown in NO.1, the sequence of the downstream primer is shown in SEQ ID NO.2, the probe is shown in SEQ ID NO.3, the 5' end of the probe sequence is modified with a fluorescent group, A substance modified with a fluorescent quenching group at the 3' end; the fluorescent group is FAM, and the fluorescent quenching group is BHQ1.

[0048] Specifically:

[0049] upstream primer wxya -F1, SEQ ID No. 1: 5'-CGCCTCATGCTCATACCC-3';

[0050] downstream primer wxya -R1, SEQ ID No.2: 5'-GAGACGCTACTGCTACCAT-3';

[0051] probe wxya -P1, SEQ ID No. 3: 5'-FAM-AATCAACCACCACACGCTCT-BHQ1-3'.

Embodiment 2

[0053] A combination of primers and probes for detecting aflatoxin-producing fungi in food, said aflatoxin-producing fungi include Aspergillus flavus and Aspergillus parasitica, said primers include upstream primers and downstream primers, and the nucleotide sequence of said upstream primers is as SEQID As shown in NO.4, the sequence of the downstream primer is shown in SEQ ID NO.5, the probe is shown in SEQ ID NO.6, the 5' end of the probe sequence is modified with a fluorescent group, A substance modified with a fluorescent quenching group at the 3' end, the fluorescent group is FAM, and the fluorescent quenching group is BHQ1.

[0054] Specifically:

[0055] upstream primer omt-1 -F2, SEQ ID No.4: 5'-CACCGAAGCCACGAAG-3';

[0056] downstream primer omt-1 -R2, SEQ ID No.5: 5'-GCTGAGATGACCACGAC-3';

[0057] probe omt-1 -P2, SEQ ID No. 6: 5'-FAM-CCGCCGACATCGACAAC-BHQ1-3'.

Embodiment 3

[0059] A combination of primers and probes for detecting aflatoxin-producing fungi in food, said aflatoxin-producing fungi include Aspergillus flavus and Aspergillus parasitica, said primers include upstream primers and downstream primers, and the nucleotide sequence of said upstream primers is as SEQID As shown in NO.7, the sequence of the downstream primer is shown in SEQ ID NO.8, the probe is shown in SEQ ID NO.9, the 5' end of the probe sequence is modified with a fluorescent group, A substance modified with a fluorescent quenching group at the 3' end, the fluorescent group is FAM, and the fluorescent quenching group is BHQ1.

[0060] Specifically:

[0061] upstream primer ver-1 -F2, SEQ ID No. 7: 5'-CCCAAACATGCCGTATACTCCGGGT-3';

[0062] downstream primer ver-1 -R2, SEQ ID No. 8: 5'-CTCCAGGAGCCACCGCATT-3';

[0063] probe ver-1 -P2, SEQ ID No. 9: 5'-FAM-CCATGCAGCGAACAAAGGTGTCA-BHQ1-3'.

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Abstract

The invention provides a primer probe combination and a kit for detecting aflatoxin-producing fungi in food, the aflatoxin-producing fungi comprise aspergillus flavus and aspergillus parasiticus, the primer probe combination is characterized in that the primer comprises an upstream primer and a downstream primer, the nucleotide sequence of the upstream primer is shown as SEQ ID NO.1, the sequence of the downstream primer is shown as SEQ ID NO.2, the probe is shown as SEQ ID NO.3, the 5' end of the probe sequence is modified with a fluorophore, and the 3' end of the probe sequence is modified with a fluorescence quenching group. The fluorophore is FAM, and the fluorescence quenching group is BHQ1. According to the primer probe combination and the kit provided by the invention, the test method is high in specificity, high in sensitivity, rapid and environment-friendly, and is suitable for rapid detection of main toxin-producing fungi in food in basic laboratories.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a combination of primers and probes and a kit for detecting aflatoxin-producing fungi in food. Background technique [0002] Fungi are widely distributed in all levels of the food chain in nature, with a wide variety and a large number, and are closely related to humans. Under suitable temperature and humidity conditions, some toxin-producing fungi will appear in food (raw materials and processed products), grain and feed grows and reproduces and produces mycotoxins. Mycotoxins are ubiquitous in agricultural by-products or animal feed, and can cause acute or chronic poisoning of humans and animals after ingestion, damage the liver, kidney, nerve tissue, hematopoietic tissue, and skin tissue of the body, and cause teratogenic and carcinogenic effects. [0003] Aflatoxin (AFT) is a highly toxic and carcinogenic mycotoxin that can easily contaminate vari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2600/166C12Q2563/107C12Q2545/113C12Q2561/113
Inventor 冯家望姚丽锋黎财慧丁琦张晴阳周昱晨游淑珠符家忍王小玉
Owner GONGBEI CUSTOMS TECH CENT
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