Kit with RT-LAMP nucleic acid test strips for detecting porcine epidemic diarrhea virus and applications of kit
A technology for RT-LAMP and porcine epidemic diarrhea, which is applied in the field of RT-LAMP nucleic acid test strip kits, can solve the problem of unstable calcein and manganese ion concentration ratio system, inaccurate discrimination of weak positive results, and limitations of application and promotion. and other problems, to achieve the effect of intuitive and accurate reaction results, low cost of use, and high sensitivity
Inactive Publication Date: 2013-09-04
SOUTH CHINA AGRI UNIV
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Problems solved by technology
However, the application of this method is limited due to the shortcomings of high false positive rate, inaccurate discrimination of weak positive results, and unstable concentration ratio system of calcein and manganese ions.
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[0059] 1. Primer design
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Abstract
The invention discloses a kit with RT-LAMP nucleic acid test strips for detecting a porcine epidemic diarrhea virus and applications of the kit. The kit comprises a primer group of a nucleic acid represented by SEQ ID No. 1-6 and test strips for detection of nucleic acid. The usage of the kit is as follows: first preparing a RT-LAMP reaction system comprising an AMV retrovirus, a 1* reaction buffer, a strand displacement DNA polymerase, a dNTP mixture, betaine, MgSO4, a FIP primer, a BIP primer, a LoopF primer, a LoopB primer, a F3 primer, a B3 primer and RNA of a sample to be measured; carrying out a reaction at a constant temperature, after testing the obtained products by using the nucleic-acid-detecting test strip, judging and reading directly: the positive result is that two red strips appear, and one strip is in the detection zone while the other strip is in the control zone. The kit has advantages of simple operation, low cost, easy observation of the reaction result, good specificity, easy popularization and application in large scope and being extremely suitable for export quarantine, food hygiene and on-site detection in animal husbandry.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to an RT-LAMP nucleic acid test strip kit for detecting porcine epidemic diarrhea virus and its application. Background technique [0002] Porcine epidemic diarrhea (porcine epidemic diarrhea, PED) is an acute, highly transmissible intestinal disease of pigs caused by porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV). The disease is characterized by acute enteritis, vomiting, watery diarrhea, and dehydration. Pigs of all ages are susceptible, especially nursing piglets, with a morbidity rate of 100% and a mortality rate of 80-100%. [0003] In 1971, porcine epidemic diarrhea was first discovered in fattening pigs and shelf pigs in the UK. In 1978, it was first identified in Belgium and the United Kingdom. Since then, except the Americas, reports of porcine epidemic diarrhea have appeared in many pig-raising countries and regions in Europe. Afterwards, many pig-...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 陈金顶勾红潮邓洁汝裴晶晶王佳莹刘文俊郑仲华
Owner SOUTH CHINA AGRI UNIV
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