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Multiple RT-PCR (reverse transcription-polymerase chain reaction) detection primer group and kit for rapidly distinguishing PEDV (porcine epidemic diarrhea virus), PDCoV (porcine deltacoronavirus) and PReoV (porcine reovirus)

A technology of RT-PCR and detection kit, which is applied in the field of animal virology and molecular biology, can solve problems such as difficulties in clinical diagnosis and detection, and difficulty in prevention and control of porcine diarrheal diseases, so as to save diagnostic reagents, shorten diagnosis time, and reduce The effect of cost of use

Inactive Publication Date: 2019-03-08
GUANGXI VETERINARY RES INST
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  • Description
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Problems solved by technology

Moreover, PDCoV is often mixed with other diarrhea pathogens, which makes clinical diagnosis and detection very difficult, and brings great difficulties to the prevention and control of porcine diarrhea diseases.

Method used

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  • Multiple RT-PCR (reverse transcription-polymerase chain reaction) detection primer group and kit for rapidly distinguishing PEDV (porcine epidemic diarrhea virus), PDCoV (porcine deltacoronavirus) and PReoV (porcine reovirus)
  • Multiple RT-PCR (reverse transcription-polymerase chain reaction) detection primer group and kit for rapidly distinguishing PEDV (porcine epidemic diarrhea virus), PDCoV (porcine deltacoronavirus) and PReoV (porcine reovirus)
  • Multiple RT-PCR (reverse transcription-polymerase chain reaction) detection primer group and kit for rapidly distinguishing PEDV (porcine epidemic diarrhea virus), PDCoV (porcine deltacoronavirus) and PReoV (porcine reovirus)

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Experimental program
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Effect test

Embodiment

[0033] 1 Materials and methods

[0034] 1.1 Viruses and clinical samples

[0035] Porcine epidemic diarrhea virus (PEDV), porcine tetracoronavirus (PDCoV), porcine reovirus (PReoV), porcine transmissible gastroenteritis virus (TGEV), swine fever virus (CSFV), porcine reproductive and respiratory syndrome Porcine encephalitis virus (PRRSV), porcine encephalitis virus (JEV), porcine bocavirus (PBoV), porcine parvovirus (PPV), porcine pseudorabies virus (PRV), porcine circovirus type 2 (PCV2) and porcine The fine ring virus (TTV) is provided by our laboratory. Clinical samples were collected from pig farms in Guangxi with diarrhea and piglet intestinal samples. About 0.5 g-1.0 g of tissue samples were cut, placed in a sterilized mortar, fully ground into a homogenate, and sterilized 1 mol / L PBS solution to make a 1:5 emulsion suspension, repeated freezing and thawing 3 times. Centrifuge at 10 000 r / min for 5 min, collect the supernatant, transfer to a 1.5 mL centrifuge tube a...

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Abstract

The invention discloses a multiple RT-PCR (reverse transcription-polymerase chain reaction) detection primer group and kit for rapidly distinguishing PEDV (porcine epidemic diarrhea virus), PDCoV (porcine deltacoronavirus) and PReoV (porcine reovirus), and belongs to the technical field of animal virology and molecular biology. The kit comprises primer sequences as shown in SEQ ID 1-6, Prime STARHR buffer solution, a cDNA template and sterile water. The multiple RT-PCR detection kit of the PEDV, the PDCoV and the PReoV is amplified by multiple RT-PCR, three lesions are similar and serve as primary detection of RNA viruses, total detection time is controlled to be about 2 hours, the detection method is easy to operate, convenient and rapid, and rapid detection can be achieved.

Description

technical field [0001] The invention belongs to the technical field of animal virology and molecular biology, and specifically relates to a detection primer set, a detection kit and application of multiple RT-PCR for PEDV, PDCoV and PReoV. Background technique [0002] Porcine epidemic diarrhea (PED) is an acute, highly contagious enteric disease caused by porcine epidemic diarrhea virus (PEDV), characterized by acute watery diarrhea, vomiting and rapid death from dehydration. Pigs of all ages can be infected, especially suckling piglets within 1-2 weeks of age are the main disease group, and the infection morbidity and mortality can even be as high as 100%. In the 1970s, Britain and Belgium respectively first reported the occurrence of the disease, and then the disease broke out and became popular in many major pig-raising countries around the world. In April 2013, the PED epidemic occurred in the United States and quickly spread to the whole country. By the end of 2014, a...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2537/143
Inventor 赵武李斌秦毅斌卢冰霞陈忠伟闭炳芬刘磊周英宁段群棚梁家幸苏乾莲何颖蒋冬福卢敬专
Owner GUANGXI VETERINARY RES INST
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