Method for identifying four fusaria

A technology of Fusarium and Fusarium oxysporum, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problems affecting the timeliness of food safety monitoring, the inability to identify pathogens in time, and the aggravation of patients' conditions and other problems, to achieve the effect of reducing the cost of experimental detection, low cost and high sensitivity

Inactive Publication Date: 2019-04-19
西安博睿康宁生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the above methods are commonly used, they take a long time, may delay treatment, make the patient's condition worse, or fail to identify the pathogen in time and affect the timeliness of food safety monitoring

Method used

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  • Method for identifying four fusaria
  • Method for identifying four fusaria
  • Method for identifying four fusaria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Differentiate the establishment and characteristic verification of four kinds of Fusarium methods

[0028] The present invention has carefully studied the genomic sequences of Fusarium proliferatum (Fusarium proliferatum), Fusarium oxysporum Schl., Fusarium semimitectum, and Fusarium solani. Look for differential target sequences in the ITS sequence, and design a pair of specific primers through careful design and screening, the nucleotide sequence of which is shown in SEQ ID NO.1-2, using this pair of primers can specifically identify the above-mentioned Four species of Fusarium. The design of this primer cannot be automatically obtained by computer software. It is necessary to select the target sequence fragment and length based on rich experience, and calculate the Tm value according to the following formula while considering the GC content of the sequence. After the primers were synthesized, trial and error was carried out to obtain a pair of optimal p...

Embodiment 2

[0046] Embodiment 2 Application of the inventive method

[0047] 1. Select 30 specimens of moldy corn kernels from Ningxia, inoculate them on SDA medium, culture them at 28°C and observe the growth of the colonies. After the growth of colonies, use the dilution coating plate method to select monoclonal colonies and culture them separately.

[0048] 2. The genomic DNA of the fungal sample to be tested was extracted by the rough extraction method, and the subsequent identification method was carried out with reference to the identification method in Example 1. The results were analyzed using ABI software, and the ITS differences of different Fusarium species were displayed based on the curve offset and curve shape changes through the standard curve. The identification results are shown in Table 2 below.

[0049] Table 2

[0050]

[0051]

[0052] For the samples for which the corresponding Fusarium was identified by the method in Example 1, the same traditional PCR meth...

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Abstract

The invention provides a method for identifying four fusaria, and belongs to the technical field of analysis of high-resolution melting curves. According to the method, a pair of specific primers areadopted, fusarium proliferatum, fusarium oxysporum, fusarium semitectum and fusarium solani are identified based on a real-time fluorescence PCR (RT-PCR) melting curve, and the sequence of the specific primers is shown in SEQ ID NO.1-2. The method is high in sensitivity, good in specificity and high in detection speed and can be used for identification of fusarium strains in the aspects of clinical diagnosis, agricultural production, food safety and the like, and therefore a reliable basis is provided for monitoring of fusaria in the aspects of infection treatment, agricultural production andfood safety.

Description

technical field [0001] The present invention relates to the technical field of multiple fungi detection, in particular to a method for distinguishing four types of Fusarium, the four types of Fusarium being Fusarium proliferatum, Fusarium oxysporum Schl., Fusarium oxysporum Schl., Fusarium semimitectum, Fusarium solani. Background technique [0002] Fusarium is a mitotic spore fungus, widely present in soil and on plants, and is an opportunistic pathogen. There are many species of Fusarium that are harmful to economic plants. They are parasites of the plant vascular system. Under suitable environmental conditions, they not only destroy the vascular bundles of crops, but also produce toxins during the growth and development of the bacteria. Harm crops, cause crop wilting, ear rot, rot, and death, affect yield and quality, and cause a significant drop in yield in severe cases. Pathogenic Fusarium oxysporum infects plants and causes plant wilt, leading to vascular bundle dise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2563/107C12Q2527/107C12Q2521/107
Inventor 周梦诗李睿蔡虎
Owner 西安博睿康宁生物科技有限公司
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