RPA (recombinase polymerase amplification) detection primer, probe and detection method for bacterial fruit blotch of cucurbit

Abstract

The invention relates to an RPA detection primer, a probe and a detection method of cucurbit bacterial fruit blotch, the RPA detection primer comprises an RPA primer pair Ac-F1 and Ac-R1 of cucurbit bacterial fruit blotch and a detection probe Ac-nfo, the amplification product size of the RPA primer pair Ac-F1 and Ac-R1 is 342bp, the 5'terminal of the primer Ac-R1 is labeled with biotin, the 5 'terminal of the primer Ac-R1 is labeled with biotin, and the 5' terminal of the detection probe Ac-nfo is labeled with biotin. The 5'tail end of the detection probe Ac-nfo is marked with FAM, the 3 'tail end of the detection probe Ac-nfo is marked with C3Spacer, and the position, 32 bp away from the 5' tail end, of the probe is modified with dSpacer; the RPA primer group is high in detection sensitivity, and the detection limit reaches 2.5 * 10 < 4 > cfu / mL; the invention provides a detection method for detecting the melon bacterial fruit blotch based on RPA combined with lateral flow test paper, establishes a rapid isothermal visual detection method for the melon bacterial fruit blotch, can complete sample detection within 15-20 min, realizes rapid diagnosis, identification and detection of the melon bacterial fruit blotch in a constant-temperature environment, and has a wide application prospect. The kit has the advantages of high sensitivity, good specificity and the like, and provides technical support for quarantine, detection, prevention and control of the disease.

Description

technical field

[0001] The invention belongs to the technical field of RPA detection, and in particular relates to a RPA detection primer, probe and detection method of melon bacterial fruit spot pathogen. Background technique

[0002] Bacterial fruit blotch (BFB) of melons is caused by Acidovorax citrulli (A.citrulli for short), which seriously damages watermelon, melon and other Cucurbitaceous crops, and is an important bacterial quarantine disease. . The disease is a disease that can be transmitted by seeds. The leaves, fruits and seeds in the growth period can be infected, which greatly affects the yield and quality of fruits, and can cause devastating damage in severe cases. In the 1990s, it was first reported in my country, and then there were occasional reports in Xinjiang, Hainan, Shanghai, Zhejiang and other places. In the "National List of Agricultural Plant Quarantine Pests Occurring in Various Regions (2016)", my country has already had 12 occurred in provinces ...

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