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Primers for detecting variants on porcine epidemic diarrhea virus and detection kit thereof

A porcine epidemic diarrhea and virus mutation technology, applied to the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve laborious, time-consuming, and high-cost problems

Inactive Publication Date: 2013-04-24
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

[0003] From the winter of 2010 to the spring of 2011, serious infectious diarrheal diseases occurred successively in pigs in pig farms all over the country. The onset was sudden, the spread was fast, the epidemic range was wide, and the epidemic time was long. The application of various antibiotics was ineffective. High mortality rate and significant economic losses
At present, other RT-PCR methods cannot solve this problem; gene sequencing technology takes a long time, and is time-consuming, laborious and costly

Method used

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  • Primers for detecting variants on porcine epidemic diarrhea virus and detection kit thereof
  • Primers for detecting variants on porcine epidemic diarrhea virus and detection kit thereof

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Embodiment 1

[0043] 1. Design and synthesis of primers

[0044] According to the nucleotide sequence of the S gene of porcine epidemic diarrhea virus mutant strain, Primer 5.0 was used to design primers, and after a large number of screenings, the primer sequences of the following sequences were obtained:

[0045] The sequence of the forward primer (PEDV-VF) is SEQ ID NO.1, and the sequence of the reverse primer (PEDV-VR) is SEQ ID NO.2. The composition of the specific sequence is as follows:

[0046] SEQ ID NO.1: 5'-TTGGTGAAAACCAGGGTGTC-3'

[0047] SEQ ID NO. 2: 5'-CAACACTATGTTCACTCA-3'.

[0048] The forward primer PEDV-VF and the reverse primer PEDV-VR were synthesized according to the above nucleotide sequence information. The above-mentioned primers PEDV-VF and PEDV-VR were respectively prepared into solutions with a concentration of 20 μmol / L for later use.

[0049] 2. A test kit for detecting variant strains of porcine epidemic diarrhea virus, assembled according to the follo...

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Abstract

The invention provides primers for detecting variants on porcine epidemic diarrhea virus and a detection kit thereof, which belongs to the technical field of biotechnology. The primers include a forward primer PEDV-VF(SEQ ID No.1) which is 5'-TTGGTGAAAACCAGGGTFTC-3' and a reverse primer PEDV-VR(SEQ ID N0.2) which is 5'-CAACACTATGTTCACTCA-3'. The invention further provides the detection kit for detecting the variants on the porcine epidemic diarrhea virus. The detection method comprises the following steps: adopting total RNA as a template, adopting the former primers to carry out inverse transcription PCR(RT-PCR) amplification, and judging according to the positions of the amplified fragments after the reaction is ended. The primer provided by the invention has good specificity. The detection method is simple and quick, has high accuracy, and provides a guarantee for detection of the variants on the porcine epidemic diarrhea virus.

Description

technical field [0001] The invention relates to a primer for detecting variant strains of porcine epidemic diarrhea virus and a detection kit thereof. It belongs to the technical field of biological detection. Background technique [0002] Porcine Epidemic Diarrhea (PED) is an acute, highly contagious infectious disease caused by Porcine Epidemic Diarrhea Virus (PEDV), often characterized by acute enteritis and dehydrated water samples Diarrhea is characteristic and pigs of all ages are susceptible. The disease was first discovered in the United Kingdom in 1971. Since then, outbreaks of the disease have also been reported in European and Asian countries. my country isolated PEDV (classic strain) in 1973. [0003] From the winter of 2010 to the spring of 2011, serious infectious diarrheal diseases occurred successively in pigs in pig farms all over the country. The onset was sudden, the spread was fast, the epidemic range was wide, and the epidemic time was long. The appli...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 李彬何孔旺杜露平郭容利倪艳秀温立斌俞正玉张雪寒茅爱华吕立新胡屹屹周俊明王小敏祝昊丹于洋
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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