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Kit for synchronously detecting six fungaltoxin producing fungi

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

Active Publication Date: 2018-07-24
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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  • Kit for synchronously detecting six fungaltoxin producing fungi

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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 kit for synchronously detecting six fungal toxins. The six toxins are aflatoxin, ochratoxin, patulin, trichothecene mycotoxin, fumonisins and zearalenone; and based on a multiple PCR detection method, the kit comprises a first primer pair, a second primer pair, a third primer pair, a fourth primer pair, a fifth primer pair, a sixth primer pair and an internal standard primer pair which can perform PCR reaction in the same reaction tube. The detection system directly targets a toxin synthesis essential gene and has high specificity; an internal reference gene is amplified synchronously, positive, negative and blank control are set simultaneously, and false negative and false positive are effectively avoided; and multiplication of PCR index guarantees 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 Applications(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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