A dual PCR method for rapid identification of goose parvovirus and cherry valley duck parvovirus

A technology of goose parvovirus and Cherry Valley Duck, applied in biochemical equipment and methods, microbe measurement/inspection, etc., to achieve the effect of improving specificity, specificity, and sensitivity

Inactive Publication Date: 2019-01-25
SHANDONG AGRICULTURAL UNIVERSITY
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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 parvovirus and this new type of goose parvovirus (Cherry Valley Therefore, there is an urgent need for a method with good specificity and high sensitivity to detect the mixed infection of traditional goose parvovirus and the new goose parvovirus

Method used

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  • A dual PCR method for rapid identification of goose parvovirus and cherry valley duck parvovirus
  • A dual PCR method for rapid identification of goose parvovirus and cherry valley duck parvovirus
  • A dual PCR method for rapid identification of goose parvovirus and cherry valley duck 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 SEQ 1 / SEQ 2 were designed for GPV and NGPV in the VP1 conserved regions of GPV and NGPV, respectively, and traditional goose parvovirus-specific Two kinds of GPV and NGPV virus samples were amplified by conventional methods using the specific primer SEQ3 and the specific primer SEQ4 of cherry valley duck parvovirus (new goose parvovirus).

[0027] SEQ 1: 5'-AGACTTATCAACAACCAT(C)T-3',

[0028] SEQ 2: 5'-TCACTTATTCCTGCTGTAG-3',

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

[0030] SEQ 4: 5'-CATCATCCGTAAAAACTTGG-3';

[0031] The amplified fragment size of the general primer SEQ 1 / SEQ 2 of GPV and NGPV is 779bp, the amplified fragment size of GPV-specific primer SEQ2 / SEQ 3 is 580bp, and the amplified fragment size of NGPV-specific primer SEQ 1 / SEQ 4 is 147bp . All primers were sterile ddH 2 O was made into a concentration of 25 μM ...

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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 (Goose parvovirus, GPV) is a member of the Parvoviridae family. It can cause disease in goslings and muscovy ducks. It produces 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 death. Rate. 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686
CPCC12Q1/6848C12Q1/701C12Q2600/16C12Q2537/143C12Q2531/113
Inventor 姜世金李鹏飞张瑞华
Owner SHANDONG AGRICULTURAL UNIVERSITY
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