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Primers for detection of melon bacterial fruit spot pathogen lamp

A fruit spot fungus, bacterial technology, applied in the biological field, can solve the problems of low specificity and low sensitivity

Inactive Publication Date: 2015-11-25
上海市农业技术推广服务中心
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are many research reports (MinsavageGVetal, 1995; Walcottetal, 2000; SchaadNWetal, 2003; SongWYetal, 2003; Baharetal, 2008) on the specific detection of Acidophage watermelon at home and abroad, but there are problems such as low sensitivity or specificity. Not strong and other shortcomings

Method used

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  • Primers for detection of melon bacterial fruit spot pathogen lamp
  • Primers for detection of melon bacterial fruit spot pathogen lamp
  • Primers for detection of melon bacterial fruit spot pathogen lamp

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Embodiment Construction

[0050] 1 Primer design

[0051] According to the sequences of Aave_4063 and Aave_4064, the Bioedit software was used for sequence analysis, and the PrimerExploreV4 software was used to design LAMP primers according to the design principles of LAMP primers. The probe was synthesized by Bao Biological Engineering (Dalian) Co., Ltd., using ddH 2 After the O was dissolved, it was aliquoted and stored in a -20°C refrigerator for later use.

[0052] 2. Establishment of LAMP reaction system

[0053] The LAMP detection reaction system is 25 μL, including 2.5 μL 10×ThermoPol (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH 4 ) 2 SO 4 , 2mMMgSO 4 , 0.1% TritionX-100, pH8.8), 8mM MgSO 4 , 0.8MBetaine, 500μMeachdNTP, 0.4μM outer primer Ac-F 3 and Ac-B 3 , 1.6μM internal primers Ac-FIP and Ac-BIP, 1μL BstDNAPolymerase (8U / μL), 1μL LoopampFlorescentDetectionReagent, 1μL DNA template or bacterial suspension, add ddH 2 O to make up to 25 μL. Place the reaction tube in a water bath at a cons...

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Abstract

The invention relates to a bacterial fruit acidovorax citrulli LAMP detection primer, belonging to the field of biotechnology. According to the whole genome sequence information of the bacterial fruit blotch strain AAC00-1, a set of specific primer for fruit blotch LAMP detection can be designed on Aave-4063 and Aave-4064 gene sequences. An LAMP detection method established on the basis of the primer has the characteristics of strong specificity, high sensitivity, low cost and the like, and is fast and easy to operate.

Description

(1) Technical field [0001] The invention relates to a LAMP primer and a detection method for melon bacterial fruit spot pathogen (Acidovorax citrulli) and belongs to the field of biotechnology. (2) Background technology [0002] Bacterial fruitblotch (BFB) of melons is an important bacterial disease on Cucurbitaceae crops such as melon, which is caused by Acidovorax citrulli. This disease is a typical seed-borne disease (SchaadNW, 1978). After the onset, it infects leaves, fruits and seeds, seriously affects the quality and yield of fruits, and poses a great threat to melon production and related industries (Richard Latin, 1995; WalcottRR, 2003), was first reported in my country in the 1990s, and then occurred in Xinjiang, Hainan, Inner Mongolia, Ningxia, Beijing, Hebei, Shandong, Jilin, Fujian, Hunan and other places, and showed an upward trend, resulting in the reduction of field watermelon and melon production Even crop failure brought huge losses to melon production (Cai ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/01
CPCC12Q1/04C12Q1/6844C12Q2531/119
Inventor 赵玉强朱建华朱恩罗金燕陈磊姚红梅余慧胡白石
Owner 上海市农业技术推广服务中心
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