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Quintuple RT-PCR detection method for the pathogen of porcine viral diarrhea

A technology of RT-PCR and porcine viral diarrhea, which is applied in the field of five-fold RT-PCR detection technology for porcine viral diarrhea pathogens, can solve problems such as poor results, and achieve reduced workload, good specificity and repeatability , The effect of improving work efficiency

Active Publication Date: 2022-04-01
华派生物技术(集团)股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Technical problem to be solved: This application mainly proposes a five-fold RT-PCR detection method for the pathogen of porcine viral diarrhea, and solves technical problems such as the poor effect of multiple RT-PCR technology in the diagnosis of viral diarrhea in pigs

Method used

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  • Quintuple RT-PCR detection method for the pathogen of porcine viral diarrhea
  • Quintuple RT-PCR detection method for the pathogen of porcine viral diarrhea
  • Quintuple RT-PCR detection method for the pathogen of porcine viral diarrhea

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Embodiment 1

[0044] A five-fold RT-PCR detection method for the pathogen of porcine viral diarrhea,

[0045] In the first step, design primer pairs for specific RT-PCR detection of 5 viruses:

[0046] Primer pair sequences for detection of porcine deltacoronavirus (Porcine Deltacoronavirus, PDCoV):

[0047] The primer sequence of the upstream primer PD-Detection (F) is 5'-TACAACCTAAGGCTAACCAAC-3',

[0048] The primer sequence of the downstream primer PD - Detection (R) is 5'- ACAACTTTACCTGCCTTACATA - 3';

[0049] Primer pair sequences for detection of Porcine Torovirus (PToV):

[0050] The primer sequence of the upstream primer PT-Detection (F) is 5'-TCCTCGTGACACTTTATCTCA-3',

[0051] The primer sequence of the downstream primer PT-Detection (R) is 5'-TTCACTAACATCCTCTACCCACA-3';

[0052] The sequence of the primer pair used to detect porcine epidemic diarrhea virus (Porcine Epidemic Diarrhea Virus, PEDV):

[0053] The primer sequence of the upstream primer PE - Detection (F) is 5'- TG...

Embodiment 2

[0069] The five-fold RT-PCR detection method of porcine viral diarrhea disease pathogen comprises the following steps:

[0070] In the first step, design primer pairs for specific RT-PCR detection of 5 viruses:

[0071] Primer pair sequences for detection of porcine deltacoronavirus (Porcine Deltacoronavirus, PDCoV):

[0072] The primer sequence of the upstream primer PD-Detection (F) is 5'-TACAACCTAAGGCTAACCAAC-3',

[0073] The primer sequence of the downstream primer PD - Detection (R) is 5'- ACAACTTTACCTGCCTTACATA - 3';

[0074] Primer pair sequences for detection of Porcine Torovirus (PToV):

[0075] The primer sequence of the upstream primer PT-Detection (F) is 5'-TCCTCGTGACACTTTATCTCA-3',

[0076]The primer sequence of the downstream primer PT-Detection (R) is 5'-TTCACTAACATCCTCTACCCACA-3';

[0077] The sequence of the primer pair used to detect porcine epidemic diarrhea virus (Porcine Epidemic Diarrhea Virus, PEDV):

[0078] The primer sequence of the upstream prim...

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Abstract

This application discloses a five-fold RT-PCR detection method for the pathogen of porcine viral diarrhea disease, in which the five pairs of primers are the detection primers of porcine delta coronavirus (also known as porcine D-coronavirus) and the detection primers of porcine protuberance virus , Porcine epidemic diarrhea virus detection primers, porcine acute diarrhea syndrome coronavirus detection primers, transmissible gastroenteritis virus detection primers. Using the nucleic acid or reverse transcription product of the sample to be tested as a template, use 2×Taq Plus Master Mix to perform PCR amplification using 5 pairs of primers in this application. The patent allows simultaneous detection of up to five viruses that can cause diarrhea in pigs in one PCR reaction tube, and its detection sensitivity reaches an initial template concentration of 1×10 per microliter 3 copies (1×10 3 copies / μL), and has excellent specificity and repeatability, and can be directly applied to the detection of clinical samples.

Description

technical field [0001] The invention relates to the technical field of pathogen detection, in particular to a five-fold RT-PCR detection technology for the pathogen of porcine viral diarrhea. Background technique [0002] In recent years, the morbidity and mortality of viral diarrhea diseases in swine herds in China and even in the world have remained high, which has brought huge economic losses to the pig breeding industry. In various parts of my country, there are frequent reports of multiple porcine diarrhea viruses mixed infection in pig herds. Mixed infections often mislead clinical veterinary workers in the diagnosis and analysis of the epidemic situation of the disease. Therefore, it is urgent to strengthen the prevention and control of highly pathogenic and infectious porcine viral diarrhea diseases including PDCoV, PEDV, PToV, Sads-CoV, and TGEV. Bearing the brunt, it is extremely necessary to establish a convenient, fast and economical detection method for porcine...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 粟硕张骋潘中洲贺微翟晓凤
Owner 华派生物技术(集团)股份有限公司
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