Real time fluorescence reverse transcription polyase chain reaction solid phase reagent kit with one-step method of citrus decline virus and testing method thereof

A decay virus and chain reaction technology, applied in the field of agricultural biology, can solve problems such as the accurate detection of citrus decay virus and the poor specificity of serology kits

Inactive Publication Date: 2008-09-24
CHONGQING UNIV
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Problems solved by technology

At present, there is no domestic production of citrus decay virus molecular diagnostic kits, and a small number of import

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment

[0055] One, the preparation of citrus decay virus harmless positive control substance: described harmless citrus decay virus positive control substance is made by the following steps:

[0056] (1) extract the total RNA containing citrus decay virus;

[0057] (2) Design the upstream and downstream primers for amplifying citrus decay virus. The upstream primer is characterized by a T7 promoter sequence at the 5' end: 5'-GCAGCTAATACGACTCACTATAGGAACAGACCATGTATAGAGGCGAAACTGCGAAT-3', and the downstream primer sequence is: 5'-TATAGAGGCGAAACTGCGAAT-3' ;

[0058] (3) carry out RT-PCR amplification to obtain the dna fragment of citrus decay virus;

[0059] (4) The RNA fragment of Citrus decay virus was transcribed in vitro by using the DNA fragment of Citrus decay virus as a template, which was used as a harmless positive control.

[0060] 2. The components of the solid-phase detection kit:

[0061] Sample viral RNA extraction reagent, viral RNA fluorescence quantitative reverse tran...

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Abstract

The invention relates to a solidification examination reagent box of one-step real-time fluorescence quantitative reverse transcription polymerase chain reaction for Citrus Tristeza Virus and a method of utilizing the reagent box to detect Citrus Tristeza Virus, which is a fluorescence quantitative detection method and product of especially establishing for Citrus Tristeza Virus. The detection time only need 4 hours and the method has the high specificity, the high sensitivity and the stability and reliability. The method overcomes the shortcomings of the high erroneous recognition and the low sensitivity of serological method for Citrus Tristeza Virus of the conventional diagnosis method according to symptomatology, is suitable for the rapid diagnosis of the viral existence of Citrus seedling and viruliferous insect sector, and can be used in quality inspection of citrus seedling and identification of Citrus Tristeza Virus.

Description

technical field [0001] The invention relates to an agricultural biotechnology, in particular to a simple and rapid solid-phase nucleic acid detection kit for one-step amplification of citrus decay virus, which is suitable for use in departments such as plant protection, plant quarantine, and agricultural product quality and technical supervision. . Background technique [0002] my country's citrus cultivation area ranks first in the world, and its total output ranks second in the world; the citrus industry has become the most important pillar industry in the three major citrus production belts in southern my country. Citrus tristeza virus (English name Citrus tristeza virus, referred to as CTV) caused by Citrus tristeza virus is an important plant disease worldwide, mainly spread by grafting and aphids, and its main hazard is to cause rapid death or serious damage to citrus plants. Citrus, which is rootstocked with limes, has been the hardest hit by the recession. The xyle...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/10
Inventor 王中康殷幼平夏玉先李正国彭国雄刘红光
Owner CHONGQING UNIV
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