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Synchronous detection kit of fungi capable of producing trichothecene mycotoxins, fumonisins, and zearalenone

A technology of trichothecenes and zearalenone, which is applied in the biological field of detection, can solve problems such as no simultaneous detection, and achieve the effects of avoiding false negatives and false positives, having strong specificity and saving labor costs.

Pending Publication Date: 2018-07-31
厦门市产品质量监督检验院(国家半导体发光器件应用产品质量监督检验中心、国家场(厂)内机动车辆质量监督检验中心)
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Problems solved by technology

Although trichothecenes, fumonisins, and zearalenone are serious hazards, current research at home and abroad focuses on the high-precision detection of the toxin itself, and there is still no simultaneous detection of bacteria producing the above three types of toxins. Multiplex PCR

Method used

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  • Synchronous detection kit of fungi capable of producing trichothecene mycotoxins, fumonisins, and zearalenone
  • Synchronous detection kit of fungi capable of producing trichothecene mycotoxins, fumonisins, and zearalenone
  • Synchronous detection kit of fungi capable of producing trichothecene mycotoxins, fumonisins, and zearalenone

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Embodiment 1

[0031] 1. Extraction of fungal genome

[0032] Toxicity-positive bacteria confirmed by HPLC (TCTs and ZEA-producing Fusarium flavum ATCC 204257 and FUM-producing Fusarium moniliform ATCC 204499, such as Figure 1 to Figure 3 shown) as a positive control; a strain that does not produce the above three types of toxins (Aspergillus flavus CGMCC 3.4408) as a negative control.

[0033] 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. Add an e...

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Abstract

The invention discloses a synchronous detection kit of fungi capable of producing trichothecene mycotoxins, fumonisins, and zearalenone. The synchronous detection kit of fungi capable of producing trichothecene mycotoxins, fumonisins, and zearalenone is invented based multiple PCR detection, and comprises a first primer pair, a second primer pair, a third primer pair, and an internal standard primer pair which are capable of realizing PCR reaction in a same reaction tube. A detection system disclosed in the invention is capable of realizing direct targeting at toxin synthesis essential genes,and is high in specificity; synchronous amplification of reference genes is realized, a positive control, a negative control, and a blank control are provided at the same time, so that false negativeand false positive are avoided effectively; and PCR index multiplication is capable of ensuring method sensitivity.

Description

technical field [0001] The invention belongs to the technical field of detection biology, and in particular relates to a simultaneous detection kit for fungi producing trichothecenes, fumonisins and zearalenone. Background technique [0002] Fungal infection generally occurs during the growth and storage periods of crops, and the biotoxins produced during the metabolic process enter the food chain of humans and animals along with the processing of grain-based food and feed, posing a serious threat to health. [0003] The genus Fusarium (Fusarium spp.) includes more than 20 species, and together with Penicillium and Aspergillus, it is the most important three types of mycotoxin-producing bacteria in nature. This genus of fungi can infect wheat, corn, oats, potatoes and other crops, among which Fusarium graminearum (F. graminearum), Fusarium sporotrichioides (F. sporotrichioides), Fusarium yellow (F. culmorum), Fusarium moniliforme ( F. proliferatum), F. verticillioides, F. o...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/16C12Q2537/143
Inventor 谢雪钦倪栋陈剑林珠白泽清
Owner 厦门市产品质量监督检验院(国家半导体发光器件应用产品质量监督检验中心、国家场(厂)内机动车辆质量监督检验中心)
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