Kit for rapidly detecting banana streak virus (BSV) by isothermal gene amplification and use method of kit

Abstract

The invention relates to a kit for rapidly detecting a banana streak virus (BSV) by isothermal gene amplification. Reagents of the kit include a primer mixed solution, a Bst DNA (deoxyribonucleic acid) polymerase with the concentration of 8U / Mu l, a 8U reverse transcriptase, an RNA extract, a reaction buffer solution, a nucleic acid dye, a positive control reagent and a negative control reagent, wherein the primer mixed solution consists of two pairs of primers; the RNA extract contains 100mM of Tris-HCL (pH 7.4), 1M of KCl, 10mM of EDTA and 2% (w / v) of PVPP; the reaction buffer solution contains 10mM of dNTP, a 10*ThermoPol reaction buffer solution, 150mM of MgSO4 and 5mM of betaine which are at a ratio of 8:5:2:10; the positive control reagent is a cDNA containing BSV coat protein; and the negative control reagent is 100mM of Tris-HCL (pH 8.0) and 50mM of EDTA. The invention also relates to a use method of the kit. By extraction of nucleic acid of the BSV from a sample and isothermal gene amplification with the kit, the color of the reaction solution is finally judged through naked eyes, so that the high-specificity, rapid, high-sensitivity, simple and convenient molecular detection on the BSV is achieved.

Description

Technical field [0001] The invention relates to the field of microbial detection. Specifically, the invention relates to a kit for rapid detection of constant temperature gene amplification of banana mosaic heart rot virus and a method of use thereof. Background technique [0002] Bananas belong to the genus Musa of the Musaceae family, and are important economic and food crops in the tropical and subtropical regions. In recent years, outbreaks of Banana Streak Virus (BSV) have occurred in major banana growing areas in my country, such as Guangdong, Yunnan, and Hainan. The early symptoms of the disease are rice-shaped chlorosis. The developmental leaves appear intermittent or continuous chlorotic strips and spindle-shaped strips, and can gradually develop into necrotic black strips. Pseudostalks, petioles and ears sometimes have striping symptoms. Bananas are mainly produced in factories through asexual tissue culture. Banana mosaic heart rot virus can reproduce with the differen...

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Problems solved by technology

[0003] In recent years, a large number of reports have confirmed that the method based on PCR has been successfully used to detect banana mosaic heart rot virus. The PCR method has greatly improved the operation time and the detection specificity and sensitivity compared with the conventional method. However, the PCR method There must be a precise temperature cycle device, and its detection time is still relatively long (2 to 3 hours), and the reaction process is easily affected by pollutants, so that this method cannot meet the needs of actual detection

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