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,

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
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Example Embodiment

[0033] Example 1. Establishment of dual PCR system and optimization of conditions

[0034] 1. Primer design and synthesis

[0035] According to the published sequence on GenBank, the DNAMan software was used to analyze the homology of PPV VP2 gene (Genbank accession number DQ464345) and PRV gE gene (Genbank accession number AF403049). According to the analysis results, Primer Primier 5.0 software was used to analyze their conserved regions. Design multiple pairs of specific primers and identify the specificity of the primers by NCBI BLAST, all of which have good specificity. The primers were synthesized by Introvigen Shanghai Biotechnology Company. The primer sequence is shown in Table 1.

[0036] Table 1 PPV and PRV double PCR primers

[0037]

[0038]

[0039] 2. Viral nucleic acid extraction

[0040] Operate according to QIAGEN kit instructions

[0041] 1) Take 20μL of proteinase K and add it to a 1.5mL sterile centrifuge tube;

[0042] 2) Add 200μL each of porcine pseudorabies viru...

Example Embodiment

[0076] Example 2. Detection of clinical samples by double PCR method

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

[0078] Tissue samples of suspected pigs from clinically affected pig farms in different regions. Please refer to the following table for sample information: including 9 pig brain tissues, 11 pig serum, 4 pig kidney and lymph node mixed samples, a total of 24 copies, and PPV is performed on the suspected samples And PRV detection. The mixed porcine pseudorabies virus PRV DNA and parvovirus PPV DNA were 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 with 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 and negative ...

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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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