A Primer Combination and a Universal Semi-Nested RT-PCR Method for the Simultaneous Identification of Three Porcine Epidemic Diarrhea Virus Strains

A technology for porcine epidemic diarrhea and epidemic diarrhea, which is applied in the field of general-purpose semi-nested RT-PCR and can solve problems such as simultaneous discrimination

Active Publication Date: 2018-09-07
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, current research reports show that three genotypes of porcine epidemic diarrhea virus strains have been discovered and identified worldwide, and there is no detection method that can distinguish these three virus strains at the same time

Method used

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  • A Primer Combination and a Universal Semi-Nested RT-PCR Method for the Simultaneous Identification of Three Porcine Epidemic Diarrhea Virus Strains
  • A Primer Combination and a Universal Semi-Nested RT-PCR Method for the Simultaneous Identification of Three Porcine Epidemic Diarrhea Virus Strains
  • A Primer Combination and a Universal Semi-Nested RT-PCR Method for the Simultaneous Identification of Three Porcine Epidemic Diarrhea Virus Strains

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Embodiment 1

[0027] Simultaneously identify the primer combination of three porcine epidemic diarrhea virus strains, the primer combination is composed of 4 specific primers, which are respectively:

[0028] Upstream primer F1: 5'-ATGGTACGTTGCTAGTGCGTA-3';

[0029] Upstream primer F2: 5'-TGGTGAAATCCAGAGTGTCAAT-3';

[0030] Upstream primer F3: 5'-CCAGCTTATATGCAGGATGGAA-3';

[0031] Downstream primer UR: 5'-TACCATGCACCAAAGTGGAATCAT-3'.

[0032] F1, F2, F3, and UR are respectively shown in sequences 1-4 in the sequence listing.

[0033] The primer design idea of ​​the present invention is as follows image 3 shown.

[0034] The universal semi-nested RT-PCR method for the simultaneous identification of three porcine epidemic diarrhea viruses using the above primer combinations comprises the following steps:

[0035] 1) Extract total RNA from the sample;

[0036] 2) Reverse transcription to obtain sample cDNA;

[0037] 3) Use the above-mentioned primers F1 and UR to carry out the first r...

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Abstract

The invention discloses a primer combination for simultaneous identification of three porcine epidemic diarrhea virus strains. The primer combination is composed of 4 specific primers: an upstream primer F1, an upstream primer F2, an upstream primer F3 and a downstream primer UR respectively. The invention also provides a general semi-nested RT-PCR method for simultaneous identification of three porcine epidemic diarrhea viruses with the primer combination. According to the primer combination and the general semi-nested RT-PCR method for simultaneous identification of three porcine epidemic diarrhea virus strains provided by the invention, all the three porcine epidemic diarrhea virus strains that are already reported and appear can be identified and distinguished simultaneously in one test by one method. The detection method is established for the first time in the world to be able to detect three different genotype porcine epidemic diarrhea virus prevalent strains at the same time, and provides a new and effective detection method for identification and clinical diagnosis of the viruses in the future.

Description

technical field [0001] The invention relates to the technical field of identification of virus strains, in particular to a combination of primers and a universal half-set RT-PCR method for simultaneously identifying three strains of porcine epidemic diarrhea virus. Background technique [0002] At present, there are many methods for porcine epidemic diarrhea virus detection at home and abroad, including ELISA, RT-PCR and fluorescent quantitative PCR, but they are all aimed at one of the original virulence strains and mutant strains of porcine epidemic diarrhea virus. Or designed and established by two strains. Therefore, current methods can only detect or differentiate strains of these two genotypes. However, current research reports show that three genotypes of porcine epidemic diarrhea virus strains have been discovered and identified worldwide, and no detection method can simultaneously distinguish these three virus strains. Contents of the invention [0003] The purp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/686
Inventor 刘新生方玉珍王永录张永光潘丽吕建亮周鹏张中旺邵军军赵付荣陈豪泰丁耀忠孙跃峰常慧芸张杰
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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