Quantitative detection method for simultaneously detecting three cat susceptible viruses and primer probe combination thereof

A quantitative detection method and primer-probe technology, which are applied in the field of quantitative detection methods for simultaneously detecting three feline susceptible viruses and primer-probe combinations thereof, can solve problems such as difficulty in quantitative detection, and achieve simple and rapid operation and specific detection. Strong and sensitive effect

Active Publication Date: 2020-01-03
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is not easy to quantify

Method used

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  • Quantitative detection method for simultaneously detecting three cat susceptible viruses and primer probe combination thereof
  • Quantitative detection method for simultaneously detecting three cat susceptible viruses and primer probe combination thereof
  • Quantitative detection method for simultaneously detecting three cat susceptible viruses and primer probe combination thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Primer design and screening:

[0054] According to the genes of each virus in GenBank, design specific probes, and label FAM, VIC or TexasRed fluorescence emitting groups at the 5' end, and label BHQ1 or BHQ2 fluorescence quenching groups at the 3' end, while targeting the probe position, For each virus, multiple pairs of primers were designed and screened, and primers with good specificity, no cross-reaction and good amplification efficiency were selected. The specific primers and probes for each virus are shown in Table 1.

[0055] Plasmid standard preparation:

[0056] Step 1: Primer Synthesis

[0057] Three pairs of viral plasmid construction primers designed by the present invention are shown in Table 2.

[0058] Step 2: Extraction of total viral nucleic acid

[0059] The DNA of FPV and FHV-1 were extracted according to the instructions of the Viral DNA Extraction Kit kit.

[0060] According to the instructions of the Viral RNA Extraction Kit kit, the RNA of FC...

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Abstract

The invention discloses a quantitative detection method for simultaneously detecting three cat susceptible viruses and a primer probe combination thereof. The sequences of the primer probe combinationare respectively FPV-F, FPV-R, FPV-P, FHV-1-F, FHV-1-R, FHV-1-P, FCV-F, FCV-R and FCV-P. The multiplex fluorescence quantitative PCR method provided by the invention can be used for simultaneously detecting three cat viruses including FPV, FHV-1 and FCV; has high detection specificity and sensitivity and simple, convenient and rapid operation, can rapidly identify and diagnose whether the cat isinfected with the virus to be detected or not in a clinical sample, can quantitatively monitor the onset condition of the cat, and provides an effective detection means for clinical monitoring of FPV,FHV-1 and FCV of the cat, epidemiological investigation, and prevention and control of the diseases.

Description

technical field [0001] The invention belongs to the technical field of quantitative detection of cat susceptibility viruses, and in particular relates to a quantitative detection method for simultaneously detecting three cat susceptibility viruses and a primer-probe combination thereof. Background technique [0002] Along with the growth of economy, the raising of people's living standard, my country's pet raising also rises day by day, has become a kind of life fashion of many people in recent years. Judging from the number of pets owned by domestic pets in China, the number of pet cats in 2018 was 67 million, accounting for 30.7% of the total number of pets, and it is still increasing year by year. At the same time, cats, as experimental animals, are more and more widely used in research fields such as medicine and biology, and the health of cats is also getting more and more attention. The three most common viruses currently affecting feline health are feline parvovirus ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/705C12Q1/701C12Q1/6851C12Q2600/16C12Q2531/113C12Q2537/143C12Q2563/107
Inventor 宋素泉闫丽萍张曦予
Owner NANJING AGRICULTURAL UNIVERSITY
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