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Kit for fluorescence quantitative PCR (polymerase chain reaction) detection of main porcine viruses

A technology of fluorescence quantification and kits, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve problems such as unsuitable for early diagnosis, low sensitivity of fluorescent antibody detection, and long time-consuming virus isolation, etc.

Active Publication Date: 2015-01-07
广州中科基因检测服务有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But in general, the above-mentioned methods have many obvious shortcomings. Virus isolation takes a long time, and the detection sensitivity of fluorescent antibodies is not very high. ELISA and serological methods can only detect antibodies, which are not suitable for early diagnosis.

Method used

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  • Kit for fluorescence quantitative PCR (polymerase chain reaction) detection of main porcine viruses
  • Kit for fluorescence quantitative PCR (polymerase chain reaction) detection of main porcine viruses
  • Kit for fluorescence quantitative PCR (polymerase chain reaction) detection of main porcine viruses

Examples

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Effect test

Embodiment 1

[0080] see figure 1 , multiple real-time fluorescent quantitative PCR detection kit, consisting of quantitative PCR reaction solution 1, porcine circovirus type 2 PCV2 standard product 2, pseudorabies virus PRV standard product 3, porcine reproductive and respiratory syndrome virus PRRSV standard product 4, classical swine fever Virus CSFV standard substance 5, porcine parvovirus PPV standard substance 6, porcine epidemic diarrhea virus PEDV standard substance 7, positive control substance 8, negative control substance 9, and box body 10.

[0081] The box body 10 is provided with container holes, respectively placed quantitative PCR reaction solution 1, porcine circovirus type 2 PCV2 standard product 2, pseudorabies virus PRV standard product 3, porcine reproductive and respiratory syndrome virus PRRSV standard product 4, classical swine fever virus CSFV Standard 5, Porcine Parvovirus PPV Standard 6, Porcine Epidemic Diarrhea Virus PEDV Standard 7, Positive Control 8, Negative...

Embodiment 2

[0083] 1 Test materials and methods

[0084] 1.1 Strains of porcine circovirus type 2, porcine parvovirus, pseudorabies virus, porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and swine fever virus.

[0085] 1.2 60 serum samples and 13 fecal samples to be tested were collected from pig farms in Fujian, Shanghai, Zhejiang and other places;

[0086] 1.3 Primers and probes

[0087] The gene sequences of porcine circovirus type 2, porcine parvovirus, pseudorabies virus, porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and swine fever virus from all over the world were downloaded from the NCBI gene bank in the United States. The homology comparison was carried out, and specific primers and TaqMan probes were designed in the conserved gene region corresponding to the virus genome. The sequence is as follows:

[0088] Upstream primer PCV virus—PR: 5'-CAGCACCCTGTAACGTTTGTC-3'

[0089] Downstream primer PCV vi...

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Abstract

The invention relates to a kit which utilizes a probe fluorescence quantitative PCR technology, is divided into two systems and can fast obtain and detect a variety of viruses causing porcine reproductive disorders and diarrheal diseases from clinical samples, wherein the A system is used for detecting porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and pseudorabies virus (PRV); the B system is used for detecting porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV) and classical swine fever virus (CSFV). The detection kit is reasonable in design, simple in a using method, fast, accurate and high in sensitivity, is suitable for port inspection and quarantine management, livestock farming, animal protection and other departments, and simultaneously has extensive scientific research values and commercial prospects.

Description

technical field [0001] The invention provides a multiple real-time fluorescence quantitative RT-PCR detection kit. technical background [0002] Every year from May to October, there will be an outbreak of a porcine reproductive disorder venereal disease characterized by high fever, dyspnea, loss of appetite, abortion and stillbirth in some sows. The disease can infect pigs of all ages. The incidence rate is high, and the mortality rate varies with the course of the disease and the severity of secondary infection fever. It is estimated that the mortality rate of severe morbidity groups can reach 50%. The disease is ferocious, and there is no specific control method, and it is easy to cause death secondary or concurrently with other pathogenic microorganisms, causing huge economic losses to the pig industry. Porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (Pseudorabiesvirus, PRV), classical swine fever virus (CSFV), p...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6851C12Q1/70C12Q2537/143C12Q2561/101C12Q2545/113
Inventor 姜永厚吴海港饶品彬
Owner 广州中科基因检测服务有限公司
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