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RT-PCR detection kit for GMBFV, PVY and GCLV viruses of garlic

A technology of RT-PCR and detection kits, applied in the direction of determination/inspection of microorganisms, DNA/RNA fragments, biochemical equipment and methods, etc.

Inactive Publication Date: 2021-05-11
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Claims
  • Application Information

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Problems solved by technology

This detection method has not been reported in the literature so far

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  • RT-PCR detection kit for GMBFV, PVY and GCLV viruses of garlic
  • RT-PCR detection kit for GMBFV, PVY and GCLV viruses of garlic
  • RT-PCR detection kit for GMBFV, PVY and GCLV viruses of garlic

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Embodiment Construction

[0023] The present invention will be further explained below in conjunction with specific examples, but the specific examples do not limit the present invention in any way.

[0024] Primer design

[0025] According to the non-conserved region of the nucleotide sequence of the viral coat protein gene included in the NCBI GenBank database, Primer5 software was used to design specific primers for the garlic mite-transmitted filamentous virus GMBFV, and the size of the amplified product was 625bp. For the primers of common latent virus GCLV, the amplified target fragment size is 450bp and the published primers for detecting potato virus Y PVY, the amplified target fragment size is 272bp, and the primer sequences are shown in Table 1. A multiplex RT-PCR detection system for GMBFV, GCLV and PVY was established. The size of the target fragments of the three viruses is quite different, there will be no overlap, and they can be clearly distinguished. The target fragments were cloned ...

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Abstract

The invention discloses a RT-PCR detection kit for GMBFV, PVY and GCLV viruses of garlic, which is characterized in that RNA of garlic tissue is used as a template, cDNA is obtained through reverse transcription, then three pairs of specific primers are adopted, one-time RT-PCR is carried out to detect the main garlic viruses simultaneously, such as the Garlic mite borne filamentous virus, the Potato virus Y and the Garlic common latent virus, and the three pairs of primers are shown in SEQ ID NO.1-SEQ ID NO.6. The detection kit provided by the invention has high detection efficiency, low detection cost and is suitable for virus detection of large-scale garlic materials.

Description

technical field [0001] The invention belongs to the technical field of crop virus detection, in particular to a garlic GMBFV, PVY, GCLV virus RT-PCR detection kit. Background technique [0002] garlic( Allium sativum L.) is an important culinary herb with an annual output of 22.28 million tons worldwide (FAO 2011) and is widely used (Keusgen M, et al, Characterization of someAllium hybrids by aroma precursors, aroma profiles , and alliinase activity. JAgric Food Chem. 2002, 50(10): 2884-90). In addition, garlic has high medicinal value (Mikaili P, et al, Therapeutic uses and pharmacological properties of garlic, shallot, and their biologically active compounds. Iran J Basic Med Sci. 2013, 16(10):1031-48) . The infection of garlic virus not only affects the yield of garlic but also affects the quality of garlic bulbs. Garlic can be infected with a variety of viruses, and usually it is a combined infection. Common viruses in garlic include Onion yellow dwarf virus (OYDV),...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/94
CPCC12Q1/701C12Q1/686C12Q2600/16
Inventor 阳文龙李锡香王海平王洋宋江萍张晓辉
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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