Duplex PCR method for rapidly identifying goose parvovirus and cherry valley duck source parvovirus

A technology of goose parvovirus and cherry valley duck is applied in the directions of biochemical equipment and methods, microorganism determination/inspection, etc., to achieve the effects of improving sensitivity, improving specificity, and improving sensitivity

Inactive Publication Date: 2016-08-03
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, PCR, fluorescent quantitative PCR, double PCR methods for detection of traditional goose parvovirus and Muscovy duck parvovirus have been established, but there is no relevant report on the detection of traditional goose parvov

Method used

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  • Duplex PCR method for rapidly identifying goose parvovirus and cherry valley duck source parvovirus
  • Duplex PCR method for rapidly identifying goose parvovirus and cherry valley duck source parvovirus
  • Duplex PCR method for rapidly identifying goose parvovirus and cherry valley duck source parvovirus

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Embodiment Construction

[0025] 1 Primer design

[0026] By comparing the sequences of GPV and NGPV published on the reference GenBank, a pair of general-specific primers SEQ1 / SEQ2 for GPV and NGPV were selected in the VP1 conserved region of GPV and NGPV, and traditional goose parvovirus-specific primers Two kinds of GPV and NGPV virus samples were amplified by conventional methods using SEQ3 and the specific primer SEQ4 of cherry valley duck parvovirus (novel goose parvovirus).

[0027] SEQ1: 5'-AGACTTATCAACAACCAT(C)T-3',

[0028] SEQ2: 5'-TCACTTATTCCTGCTGTAG-3',

[0029] SEQ3: 5'-CCGTTCCCGTCGGATGTC(G)-3',

[0030] SEQ4: 5'-CATCATCCGTAAAAACTTGG-3';

[0031] The fragment size amplified by the general primers SEQ1 / SEQ2 of GPV and NGPV is 779bp, the fragment size amplified by the specific primers SEQ2 / SEQ3 of GPV is 580bp, and the fragment size amplified by the specific primers SEQ1 / SEQ4 of NGPV is 147bp. All primers were sterile ddH 2 O was made into a concentration of 25 μM for use.

[0032] 2D...

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Abstract

The invention relates to a duplex PCR method for rapidly identifying the goose parvovirus and the cherry valley duck source parvovirus. New specific primers are designed and screened, various parameters in the reaction process are optimized, and the duplex PCR method, established with the group of primers, for the goose parvovirus and the cherry valley duck source parvovirus is high in specificity and sensibility and capable of being used for rapidly and accurately identifying the goose parvovirus and the new cherry valley duck source parvovirus. The established duplex PCR method is good in repeatability, high in reliability, low in cost, easy to implement and suitable for large-scale epidemiological investigation. It is proved through detection of 1500 samples that the duplex PCR method has high reliability; besides, identification can be carried out just through a common PCR instrument, the cost of primer synthesis is very low, and thus the cost of a reaction in the duplex PCR method is almost the same as that of a common PCR; experiment operation is also easy, and the duplex PCR method is suitable for the present production fact of the animal husbandry in China.

Description

(1) Technical field [0001] The invention relates to a double PCR method for quickly identifying goose parvovirus and cherry valley duck parvovirus, belonging to the field of virus molecular biology. (2) Background technology [0002] Goose parvovirus (Gooseparvovirus, GPV) is one of the members of Parvoviridae, which can cause disease in goslings and muscovy ducks, and produce lesions characterized by the necrosis and shedding of the surface of the small intestinal mucosa and the formation of small intestinal embolism, with high morbidity and mortality . According to previous studies, the natural infection hosts of parvovirus in waterfowl are only geese and Muscovy ducks, and there are no reports of it infecting other species of ducks. However, since 2015, there have been reports in my country that Cherry Valley meat ducks have symptoms such as unexplained short beak, protruding tongue, slow growth, and weight loss. Clinical autopsy showed enlarged thymus, obvious bleeding ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6848C12Q1/701C12Q2600/16C12Q2537/143C12Q2531/113
Inventor 姜世金李鹏飞张瑞华
Owner SHANDONG AGRICULTURAL UNIVERSITY
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