A simultaneous detection kit for six types of mycotoxin-producing bacteria

A mycotoxin and synchronous detection technology, applied in the field of biological detection, can solve the problems of time-consuming and complicated professional requirements, and achieve the effects of avoiding false negatives and false positives, ensuring sensitivity, and reducing harm

Active Publication Date: 2022-01-18
厦门市产品质量监督检验院
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Problems solved by technology

[0003] At present, the routine identification methods of toxin-producing fungi are mostly based on morphological and fertility analysis (such as GB 4789.16-2016 National Food Safety Standard Food Microbiology Examination of Morphological Identification of Common Toxigenic Fungi; SN / T 1035-2011 Imported and Penicillium, Aspergillus and their toxin detection methods), time-consuming, tedious and highly professional requirements

Method used

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  • A simultaneous detection kit for six types of mycotoxin-producing bacteria

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Embodiment 1

[0027] 1. Extraction of fungal genome

[0028] Positive bacteria that produce aflatoxins, ochratoxins, patulin, trichothecenes, fumonisins and zearalenone (A. CGMCC3.6890, Fusarium yellow ATCC 204257 and Fusarium moniliforme ATCC 204499) were used as positive controls; strains that did not produce the above six toxins (Cordyceps militaris XMZJ-F-001) were used as negative controls.

[0029] Inoculate the working bacteria (spores or hyphae) of the above-mentioned strains on a PDA plate with cellophane on the surface, culture at 25°C for 3-4 days, and harvest the mycelium when the mycelium is covered with the culture dish and no spores are produced, refer to the following method Extraction of DNA: Weigh 0.2 g of mycelium, place it in a pre-cooled mortar, and grind it to powder with liquid nitrogen. Then add 4mL DNA extraction solution [0.2M Tris·HCl (pH 7.5), 0.5M NaCl, 10mM EDTA, 1% SDS (w / v)], continue grinding, transfer to a centrifuge tube, and place in ice bath for 3-5min....

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Abstract

The invention discloses a simultaneous detection kit for fungi producing six toxins, the six toxins being aflatoxin, ochratoxin, patulin, trichothecene toxins, fumonisins and zearalen Ketone, based on the multiplex PCR detection method, including the first primer pair, the second primer pair, the third primer pair, the fourth primer pair, the fifth primer pair, the sixth primer pair and the inner indexer pair. The detection system of the present invention directly targets genes necessary for toxin synthesis, and has strong specificity; internal reference genes are synchronously amplified, and positive, negative and blank controls are set at the same time, effectively avoiding false negatives and false positives; PCR index multiplication ensures the sensitivity of the method.

Description

technical field [0001] The invention belongs to the technical field of detection biology, and in particular relates to a simultaneous detection kit for bacteria producing six types of mycotoxins. Background technique [0002] Fungal infections are common in the growth and storage periods of cereal crops. Among them, the infection of fungi of Aspergillus, Penicillium and Fusarium not only reduces grain production and quality, but also produces a series of toxins during its metabolism, which seriously endangers health. Aflatoxins, ochratoxins, patulins, trichothecenes, zearalenone and fumonisins are the six most harmful types of mycotoxins in the world. Eating or long-term exposure to primary agricultural products, feed and processed food contaminated by the above-mentioned toxins will cause body damage, organ lesions and even cancer. [0003] At present, the routine identification methods of toxin-producing fungi are mostly based on morphological and fertility analysis (suc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11C12R1/67C12R1/66C12R1/77
CPCC12Q1/6895C12Q2600/166C12Q2600/16
Inventor 谢雪钦高静倪栋陈剑林珠刘燕頔
Owner 厦门市产品质量监督检验院
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