Kit for rapidly detecting various viruses in muskmelon by one step and rapid detection method adopting kit

A melon yellow spot virus and kit technology, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microbial determination/inspection, etc., can solve the problems of few types of melon virus diseases, and achieve low equipment requirements and high accuracy. , the effect of fewer steps

Inactive Publication Date: 2016-02-17
INST OF PLANT PROTECTION OF XINJIANG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problem that the existing muskmelon virus disease multiple RT-PCR detection technology can simultaneously d

Method used

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  • Kit for rapidly detecting various viruses in muskmelon by one step and rapid detection method adopting kit
  • Kit for rapidly detecting various viruses in muskmelon by one step and rapid detection method adopting kit
  • Kit for rapidly detecting various viruses in muskmelon by one step and rapid detection method adopting kit

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specific Embodiment approach 1

[0039] Specific embodiment one: the kit for the rapid detection of multiple viruses in muskmelon contains 7 pairs of RT-PCR primers in this embodiment;

[0040] A pair of primers for amplifying watermelon mosaic virus disease: the nucleotide sequence of the upstream primer WMV1F is 5'-GAGGAACTGTGGTTCATGTC-3', and the nucleotide sequence of the downstream primer WMV1R is 5'-GCAGTGTGCCTCTCAGTATT-3';

[0041] A pair of primers for amplifying cucumber mosaic virus disease: the nucleotide sequence of the upstream primer CMV1F is 5'-GACAGTTGGGAATCGGA-3', and the nucleotide sequence of the downstream primer CMV1R is 5'-AACAGGGAGCAAGAGGA-3';

[0042] A pair of primers used to amplify the yellow mosaic virus of zucchini: the nucleotide sequence of the upstream primer ZYMV-F is 5'-CACGAAGGACAAGGATGTGA-3', and the nucleotide sequence of the downstream primer ZYMV-R is 5'-ACATTGCTAAGAGCTGCTGC-3 ';

[0043] A pair of primers for amplifying squash mosaic virus: the nucleotide sequence of t...

specific Embodiment approach 2

[0047] Specific embodiment two: present embodiment one step rapid detection muskmelon multiple virus carries out according to the following steps:

[0048] 1. Extract the total RNA of infected virus leaves;

[0049] 2. Reverse transcription into cDNA;

[0050] 3. Perform multiple RT-PCR with 7 pairs of RT-PCR primers in the kit to determine whether the leaves are infected with watermelon mosaic virus, cucumber mosaic virus, zucchini yellow mosaic virus, squash mosaic virus, Tobacco mosaic virus, papaya ringspot virus and melon yellow spot virus, and virus types;

[0051] In step 3, 7 pairs of RT-PCR primers are primer pairs for amplifying watermelon mosaic virus: the nucleotide sequence of the upstream primer WMV1F is 5'-GAGGAACTGTGGTTCATGTC-3', and the nucleotide sequence of the downstream primer WMV1R is 5 '-GCAGTGTGCCTCTCAGTATT-3';

[0052] A pair of primers for amplifying cucumber mosaic virus disease: the nucleotide sequence of the upstream primer CMV1F is 5'-GACAGTTGG...

specific Embodiment approach 3

[0060] Embodiment 3: The difference between this embodiment and Embodiment 2 is that in Step 2, random primers and reverse transcriptase are used for reverse transcription. Other steps and parameters are the same as those in Embodiment 2.

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Abstract

The invention provides a kit for rapidly detecting various viruses in a muskmelon by one step and a rapid detection method adopting the kit, relates to a kit for rapidly detecting muskmelon virus diseases and a rapid detection method adopting the kit and solves the problem that a conventional multiplex RT-PCR (reverse transcription-polymerase chain reaction) detection technology for the muskmelon virus diseases can detect a small variety of muskmelon virus diseases simultaneously. The kit for rapidly detecting the various viruses in the muskmelon by one step contains 7 pairs of RT-PCR primers. The rapid detection method comprises the following steps: 1, total RNA (ribonucleic acid) of a blade infected with viruses is extracted; 2, the total RNA is reversely transcribed into cDNA (complementary DNA); 3, multiplex RT-PCR is performed by the 7 pairs of RT-PCR primers in the kit. As many as 7 muskmelon virus diseases can be detected simultaneously through one-time PCR, and the kit and the method can be used for gathering and identifying muskmelon virus disease varieties in a field and have the advantages of simple identification process, a few steps, low equipment requirements, low cost, short time, high accuracy and the like.

Description

technical field [0001] The invention relates to a kit for rapidly detecting muskmelon virus disease and a rapid detection method thereof. Background technique [0002] Melon virus disease has always been the main factor limiting the production of melon. At present, there is no effective method to prevent and control melon virus disease at home and abroad. It is common for viral diseases to be combined with disease in melon producing areas. The methods for detecting muskmelon virus disease virus species at present have methods such as double-antibody sandwich method (DAS-ELISA) and nitrocellulose membrane method (NCM-ELISA) method, serum antigen detection; long. [0003] Multiplex RT-PCR technology can detect several pathogenic bacteria at the same time, which greatly shortens the inspection time and process, saves the test materials, and is especially suitable for the rapid detection of germplasm materials or pathogenic isolates that are difficult to separate and obtain. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/94
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2537/143C12Q2521/107
Inventor 玉山江·麦麦提杨渡韩盛李继洋丁新华吐尔逊·阿合买提何丹米日古丽·马木提
Owner INST OF PLANT PROTECTION OF XINJIANG ACADEMY OF AGRI SCI
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