PCR quick detection method for bee European foul brood

A detection method, the technology of putrefaction disease, applied in the biological field, can solve the problems of time-consuming, judgment easily affected by subjective factors, etc., achieve the effect of simple detection method, control bee disease, and improve the quality of bee products

Inactive Publication Date: 2010-08-04
吉林出入境检验检疫局检验检疫技术中心
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Problems solved by technology

This method mainly relies on morphological examination, which takes a long time and is easily affected by subjective factors.

Method used

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  • PCR quick detection method for bee European foul brood
  • PCR quick detection method for bee European foul brood
  • PCR quick detection method for bee European foul brood

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Experimental program
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Embodiment Construction

[0030] Concrete steps of the present invention are:

[0031] a. Using the CTAB method or the kit method to obtain the European larval rot disease DNA;

[0032] b. Determination of the concentration and purity of European larval rot DNA: take 5 μL of DNA solution and dilute it to 1 mL with double distilled water, and use a nucleic acid protein analyzer or an ultraviolet spectrophotometer to detect the absorbance values ​​A260 and A280 at 260nm and 280nm respectively, so that A260 The ratio to A280 is between 1.7 and 2.0;

[0033] c. Primers:

[0034] EFB1: 5'-CTT TGAACG CCT TAG AGA-3'

[0035] EFB2: 5'-ATC ATC TGT CCC ACC TTA-3'

[0036] EFB3: 5'-TTAACC TCG CGG TCT TGC GTC TCT C-3';

[0037] d. Using the semi-nested PCR amplification method, according to the 16S rRNA gene of Apis coccus hives, design primers EFB1 and EFB2 for the first round of PCR amplification. The fragment size of the obtained PCR theoretical amplification product is 486bp. The amplification result of th...

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Abstract

The invention discloses a PCR quick detection method for bee European foul brood, belongs to the field of biology and aims to provide a quick, accurate and convenient PCR quick detection method for the bee European foul brood for the diagnosis, prevention and control, quarantine and bee product disease cause contamination detection of the bee European foul brood by establishing a PCR detection method. The method has the advantages of high speed, high sensitivity, high specificity and the like, can be used for quickly diagnosing the European foul brood, and is more effective particularly for differential diagnosis of mixed infection caused by a plurality of disease causes. The PCR detection method established by the standard provides the quick, accurate and convenient PCR quick detection method for the bee European foul brood for the diagnosis, prevention and control, quarantine and bee product disease cause contamination detection of the European foul brood in China so as to effectively control bee diseases and has great significance for improving the quality of bee products.

Description

Technical field: [0001] The present invention belongs to the biological field. Background technique: [0002] Honeybee European foulbrood, referred to as European foulbrood (EFB) is a contact infectious disease in the larval stage of honeybees. It mainly infects small larvae under the age of 2 days, but when the course of the disease is prolonged, the capped larvae may also die. In this case, it is easy to be confused with the beautiful rot stick. Its pathogenic bacteria do not form spores and are easier to cure. [0003] European young disease, widely occurs in various beekeeping countries, is a worldwide disease. The disease was first reported in 1885, and now it occurs widely in almost all beekeeping producing countries in the world. It was first discovered in Guangdong Province in the early 1950s in China, and the disease appeared in the southern provinces in the early 1960s, and then spread throughout the country. The disease is not only infected by Western honeybee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/01
Inventor 宋战昀王振国罗雁非刘金华肖成蕊石建平蔡阳孟庆峰
Owner 吉林出入境检验检疫局检验检疫技术中心
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