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Kit for detecting porcine pseudorabies virus (PRV) and porcine parvovirus (PPV)

A technology for porcine pseudorabies virus and porcine pseudorabies disease, which can be applied in the biological field and can solve problems such as economic losses in the pig industry

Active Publication Date: 2011-04-13
CHINA ANIMAL DISEASE CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many pig farms have two diseases at the same time, causing great economic losses to the pig industry

Method used

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  • Kit for detecting porcine pseudorabies virus (PRV) and porcine parvovirus (PPV)
  • Kit for detecting porcine pseudorabies virus (PRV) and porcine parvovirus (PPV)
  • Kit for detecting porcine pseudorabies virus (PRV) and porcine parvovirus (PPV)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1, establishment of dual PCR system and optimization of conditions

[0034] 1. Primer design and synthesis

[0035] According to the sequence published on GenBank, DNAMan software was used to analyze the homology of PPV VP2 gene (Genbank accession number DQ464345) and PRV gE gene (Genbank accession number AF403049). Multiple pairs of specific primers were designed and identified by NCBI BLAST, all of which had good specificity. Primers were synthesized by Introvigen Shanghai Biotechnology Company. The primer sequences are listed in Table 1.

[0036] Table 1 PPV and PRV double PCR primers

[0037]

[0038]

[0039] 2. Viral nucleic acid extraction

[0040] According to the instructions of the QIAGEN kit

[0041] 1) Add 20 μL proteinase K (proteinase K) into a 1.5 mL sterilized centrifuge tube;

[0042] 2) Add 200 μL each of porcine pseudorabies virus PRV and porcine parvovirus PPV, and mix well. If the volume is less than 200 μL, make up with PBS; ...

Embodiment 2

[0076] Embodiment 2, carry out the detection of clinical sample with double PCR method

[0077] 1. Detection of PPV and PRV in clinical suspected samples

[0078] Tissue samples of suspected diseased pigs from clinically affected pig farms in different regions, please see the following table for sample information: 24 mixed samples including 9 pig brain tissues, 11 pig serum, 4 pig kidneys and lymph nodes, PPV for suspected samples and PRV detection. The mixed DNA of porcine pseudorabies virus PRV and parvovirus PPV was used as a positive control, and sterilized deionized water was used as a negative control. Perform PCR detection according to the optimized double PCR system and double PCR reaction conditions obtained in Example 1.

[0079] Test results such as Figure 7 and Figure 8 shown.

[0080] Figure 7 Clinical sample test results 1, M: Trans DNA Marker I, the size from top to bottom is: 700, 600, 500, 400, 300, 200, 100; 1, 2: positive control, negative control, r...

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Abstract

The invention discloses a kit for detecting porcine pseudorabies virus (PRV) and porcine parvovirus (PPV), which comprises a primer pair 1 and a primer pair 2, wherein one primer in the primer pair 1 is the DNA molecule disclosed as sequence 1 in the sequence table, the other primer in the primer pair 1 is the DNA molecule disclosed as sequence 2 in the sequence table, one primer in the primer pair 2 is the DNA molecule disclosed as sequence 3 in the sequence table, and the other primer in the primer pair 2 is the DNA molecule disclosed as sequence 4 in the sequence table. The experiment proves that the kit can be used for quickly, specifically and sensitively detecting PRV and PPV from the specimen, and lays foundation for clinical quick diagnosis and epidemiology research of PRV and PPV.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting porcine pseudorabies virus and porcine parvovirus. Background technique [0002] Porcine pseudorabies (PRV) and parvovirus (PPV) are common diseases in the pig industry that cause reproductive disorders in pigs, and are also associated with weaned multiple systemic failure syndrome (PMWS), porcine dermatitis nephrotic syndrome (PDNS), Porcine proliferative necrotizing pneumonia (PNP) is also closely related. PRV and PPV are often mixed infection. Among them, PRV is very harmful to pigs, and can cause abortion, stillbirth, and mummified fetuses in pregnant sows. Newborn piglets have obvious neurological symptoms, and the mortality rate is almost 100%. PPV mainly causes reproductive disorders in pigs. Pigs are characterized by stillbirths, deformed fetuses, mummified fetuses and sickly piglets. Both diseases exist in many pig farms at the same time, causing great...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 夏应菊訾占超顾小雪韩雪汪葆玥倪建强曲萍翟新验田克恭
Owner CHINA ANIMAL DISEASE CONTROL CENT
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