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Multiplex RPA detection method for lethal necrosis of corn

A detection method and technology for necrosis, which are applied in the field of multiple RPA detection of lethal necrosis of corn, can solve problems such as simultaneous detection that has not yet been found, and achieve the effects of simple RPA equipment, good specificity and short response time.

Active Publication Date: 2021-03-09
SHENYANG AGRI UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] RPA (Recombinase polymerase amplification) is a new type of DNA isothermal amplification technology, which can amplify template DNA sensitively, specifically and rapidly, and can also detect plant viruses on site. There is no report on the simultaneous detection of two plant viruses using RPA technology.

Method used

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  • Multiplex RPA detection method for lethal necrosis of corn
  • Multiplex RPA detection method for lethal necrosis of corn
  • Multiplex RPA detection method for lethal necrosis of corn

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Experimental program
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Embodiment 1

[0047] 1. Determination of specific primers for SCMV and MCMV:

[0048]Reference SCMV genome (GenBank number: KT736022.1, KR611106.1, AY042184.1, AJ310109.1, AJ271085.1, AJ310111.1, AJ310106.1, AJ421461.1, AJ421463.1AJ310107.1) and MCanMVk genome No.: MH645620.1, X14736.2, EU358605.1, MK491605.1, MN756483.1, MK684213.1, MK684203.1, MK684201.1, MF467375.1, MH205605.1) sequence, according to the design principles of RPA primers , using software and manual correction to design the specific RPA primer pairs SCMV-F / SCMV-R and MCMV-F / MCMV-R for simultaneous detection of SCMV and MCMV, the primer sequences are shown in Table 1;

[0049] The specific RPA primers of the sugarcane mosaic virus are SCMV-F, SCMV-R, and the specific RPA primers of the corn chlorotic mottle virus are MCMV-F and MCMV-R;

[0050] The primer sequence of SCMV-F is shown in SEQ ID NO.1;

[0051] 5'-CAAGATATATCAAACACTAGAGCAACTAAG-3'

[0052] The primer sequence of SCMV-R is shown in SEQ ID NO.2;

[0053] 5'-G...

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Abstract

The invention belongs to the field of plant virus detection methods and particularly relates to a multiplex RPA detection method for lethal necrosis of corn. The method comprises the steps of carryingout sample nucleic acid extraction, carrying out reverse transcription, carrying out RPA reaction specificity detection, carrying out cataphoresis, and judging whether a sample carries over two kindsof viruses, i.e., MCMV and SCMV or not. The method disclosed by the invention is good in specificity, relatively high in sensitivity and short in reaction time.

Description

technical field [0001] The invention belongs to the field of plant virus detection methods, in particular to a multiple RPA detection method for corn lethal necrosis. Background technique [0002] Sugarcane mosaic virus (SCMV) is a member of the Potyvirus genus of the family Potyviridae, and is mainly transmitted by aphids in a non-persistent manner, and can also be transmitted by sap and seeds. After SCMV infects plants, mosaic symptoms are formed on the surface of leaves, which can also cause dwarfing of plants and cause maize dwarf mosaic disease. Maize dwarf mosaic is a worldwide viral disease, and sugarcane mosaic virus Beijing isolate (SCMV-BJ) is the dominant strain causing maize dwarf mosaic in the corn-producing areas of northern my country. [0003] Maize chlorotic mottle virus (MCMV) belongs to the genus Machlomovirus of the family Tombusviridae, and is mainly transmitted by sap, vector beetles, and thrips, and can also be transmitted by seeds. After MCMV infect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12R1/94
CPCC12Q1/701C12Q1/6844C12Q2600/16C12Q2521/107C12Q2537/143C12Q2565/125
Inventor 夏子豪高新然安梦楠吴元华王志华郝凯强罗雪琮
Owner SHENYANG AGRI UNIV
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