PCR (Polymerase Chain Reaction) and fluorescent PCR rapid detection method of bee sacbrood diseases

A technology for cystic larvae and detection methods, applied in fluorescence/phosphorescence, biochemical equipment and methods, and microbial determination/inspection, etc., can solve the problems of low sensitivity and specificity, reduce drug residues, and facilitate detection method, control effect

Inactive Publication Date: 2011-01-26
中华人民共和国吉林出入境检验检疫局
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  • Abstract
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  • Application Information

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Problems solved by technology

So far, the laboratory diagnosis of cystic larval disease virus is mainly based on the observation of electron microscope, as well as traditional detection methods such as agar diffusion test, radioimmunoassay, ELISA and other immunological methods. The sensitivity and specificity of these methods Low to avoid false positive reactions

Method used

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  • PCR (Polymerase Chain Reaction) and fluorescent PCR rapid detection method of bee sacbrood diseases
  • PCR (Polymerase Chain Reaction) and fluorescent PCR rapid detection method of bee sacbrood diseases
  • PCR (Polymerase Chain Reaction) and fluorescent PCR rapid detection method of bee sacbrood diseases

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Embodiment Construction

[0029] The steps of the present invention are:

[0030] a, first collect adult bee specimens and larval specimens;

[0031] b. Primer design and synthesis:

[0032] Primers for PCR detection of bee cystic larvae:

[0033] SBV 1 5’-ACC AAC CGA TTC CTC AGT AG-3’

[0034] SBV 2 5'-CCT TGG AAC TCT GCT GTG TA-3'

[0035] Taqman probes and primers for real-time fluorescent PCR detection of bee cystic larvae:

[0036] SBV311F 5'-AAGTTGGAGGCGCGTAATTG-3'

[0037] SBV380R 5'AAATGTCTTCTTACTAGAGGTAAGGATTG-3'

[0038] SBV331T 5’-FAM-CGGAGTGGAAAGATTATCTACAATCCTTACCTCTA-TAMRA-3’

[0039] c. (1) Take a single bee, wash it with PBS solution for several times, discard as much excess PBS as possible, add 500 μL Trizol reagent, grind as much as possible with a grinder, then add 500 μL Trizol reagent, mix quickly, and stand at room temperature for 10 minutes ;

[0040] (2) Add 200 μL of chloroform, close the lid tightly, shake vigorously by hand for 15 seconds, and let stand at room ...

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) and fluorescent PCR rapid detection method of bee sacbrood diseases, belonging to the field of biology and aiming at establishing a PCR detection method and providing a rapid, accurate, simple and convenient PCR (Polymerase Chain Reaction) and fluorescent PCR rapid detection method of bee sacbrood diseases, for the diagnosis, control and quarantine of the bee sacbrood diseases and the pathogenic pollution detection of bee products. The method comprises the step of designing and synthesizing primers for PCR detection of the bee sacbrood diseases, as well as a Taqman probe and primers for real-time fluorescent PCR detection of the bee sacbrood diseases. The invention provides the rapid, accurate, simple and convenient method for the diagnosis, control and quarantine of the bee sacbrood diseases and the pathogenic pollution detection of the bee products, and provides technical guarantees for effectively controlling the propagation of the sacbrood diseases, reducing drug residue in bees and improving the quality of the bee products.

Description

Technical field: [0001] The present invention belongs to the biological field. Background technique: [0002] Sacbrood disease (sacbrood disease) is referred to as sac larvae disease for short, also known as point disease and sac brood disease. It is an intestinal infectious disease of bee larvae caused by sacbrood bee virus (SBV). one of the diseases. It happens from time to time in beekeeping production. When the disease is mild, nearly a hundred larvae die, and worker bee collection activities are normal; when the disease is severe, the bee colony only sees an increase in larvae but no increase in adult bees, resulting in the death of a large number of larvae, and worker bees are restless. The collection power dropped significantly, and even the whole group flew away, causing serious losses to the beekeepers. [0003] Bee sac larval disease was first discovered in 1913, but it was not confirmed until 1964 that its pathogen was sac larvae virus [9] , In 1971, cystic juv...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 王振国宋战昀刘和平石建平罗雁非刘金华魏春艳肖成蕊
Owner 中华人民共和国吉林出入境检验检疫局
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