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RPA primer and detection method for detecting banana streak virus GF isolate

A Banana Streak Virus, BSGFV-RPA-F technology, applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc.

Pending Publication Date: 2020-10-23
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is no report on the RPA detection technique for GF isolates of banana streak virus

Method used

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  • RPA primer and detection method for detecting banana streak virus GF isolate
  • RPA primer and detection method for detecting banana streak virus GF isolate
  • RPA primer and detection method for detecting banana streak virus GF isolate

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Experimental program
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Effect test

Embodiment

[0036] 1. Design of RPA primers and probes

[0037] 1. Design specific RPA primers based on the coat protein gene in the banana streak virus GF isolate. The coat protein gene sequence is shown in SEQ ID NO.1.

[0038] 2. The nucleotide sequence of the RPA primer is:

[0039] Upstream primer BSGFV-RPA-F: CTTGGCAAAATTGGAAGGATCTAGATTACC SEQ ID NO.2;

[0040] Downstream primer BSGFV-RPA-R: CTTAGATGATGAATTGGCTCTTCCAC SEQ ID NO.3;

[0041] The fragment size of the amplified product was 401bp.

[0042] 3. The nucleotide sequence of the probe is P:

[0043] CTGTCTTCATACTCTCGATCTTAGAACT(FAM-dT)(dSpacer)A(BHQ1-dT)AATTGCAGGAGCAGGAATTACAGCTC-C3spacer, as shown in SEQ ID NO.4;

[0044]Among them, probe P has (FAM-dt) group and (dspacer) modification added at the 28th bp position, (BHQ1-dT) group is added at the 29th bp position, and (C3spacer) modification is added at the tail .

[0045] 2. Kit composition and method of use

[0046] The RPA kit also includes the RPA reaction amplifi...

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Abstract

The invention discloses an RPA primer and a detection method for detecting a banana streak virus GF isolate, and relates to the field of biotechnology. Aiming at a conserved sequence of a coat proteingene in the banana streak virus GF isolate, the invention designs the specific RPA primer, a probe and a kit, and accurate qualitative detection can be performed on the banana streak virus GF isolate. Based on the RPA amplification primer and the probe, the kit has high accuracy and detection sensitivity of 0.1ng, and is simple and rapid to operate, provides technical support for gene specificitydetection in the banana streak virus GF isolate, has scientific guiding significance for banana virus disease prevention and control and product inspection and quarantine, and also has important significance for reducing blind use of pesticides, reducing the production cost and reducing pesticide environmental pollution.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, it relates to an RPA primer and a detection method for detecting banana stripe virus GF isolates. Background technique [0002] Banana streak virus (BSV) is one of the important viral diseases of banana. BSV belongs to the genus Badnavirus of the family Caulimoviridae, and is a circular double-stranded DNA with a genome size of 7.5-8.0 kb. Bananas infected by BSV can produce a variety of symptoms. The typical symptoms are intermittent and / or continuous chlorotic stripes and shuttle stripes on the leaves. With the development of symptoms, it can gradually become necrotic black stripes. According to the report of the Tenth International Committee on Taxonomy of Viruses (ICTV), BSV is divided into 9 species, which are Banana streak GF virus (BSGFV), Banana streak UA virus (Banana streakUA virus, BSUAV), banana Banana streak IM virus (BSIMV), Banana streak MY virus (BSMYV), Banana st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2521/507C12Q2522/101
Inventor 余乃通刘志昕李俊成高圣风
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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