Detection method of erwinia stewartii loop-mediated isothermal amplification test paper strip
A ring-mediated isothermal, fusarium wilt technology, applied in microorganism-based methods, biochemical equipment and methods, and microbial assay/inspection, etc., can solve problems such as inconvenience, misjudgment, and high cost of naked eye observation.
Inactive Publication Date: 2019-03-22
CHINA INSPECTION LAB TECH CO LTD
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Problems solved by technology
The first two judgment methods will cause defects such as inconvenient visual observation and misjudgment due to the inconspicuous result color, while the latter two require the purchase of expensive analytical instruments. Therefore, it is necessary to find a cheap and convenient and accurate observation method with the naked eye. Detection method
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[0021] 1. Test materials and methods
[0022] 1.1 Test material
[0023] Each strain was cultured and amplified under standard procedures, and bacterial genomic DNA was extracted using TIANamp bacteria DNAkit. Bacterial Fusarium wilt of corn wxya Escherichia coli with gene recombination plasmid, 160~180r, 37°C, 45° inclined culture for 14 hours. Extract the plasmid and determine the concentration.
[0024] 1.2 Primer design
[0025] Using MEGA6 software to control corn bacterial wilt wxya compare genes, select wxya The fragment of the conserved region of the gene is used as the target sequence, and the online software PrimerExplorer V5 (http: / / primerexplorer.jp / e / ) is used to design wxya Loop-mediated isothermal amplification of genes with lateral flow dipstick primer sets. figure 2 shown wxya The gene loop mediated isothermal amplification lateral flow test strip primer system consists of a pair of outer primers (SEQ No.1, SEQ No.2), a pair of inner primers (SEQ...
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The invention discloses a loop-mediated isothermal amplification transverse flowing test paper strip for detecting erwinia stewartii and an application method thereof. A detection method is characterized in that the loop-mediated isothermal amplification transverse flowing test paper strip is composed of an erwinia stewartii wtsE gene loop-mediated isothermal amplification transverse flowing testpaper strip primer mixed solution, a loop-mediated isothermal amplification reaction pre-mixed solution and positive and negative quality control products of erwinia stewartii pathogenic bacteria. A primer group is characterized by being formed by designing an erwinia stewartii wtsE gene conserved region, and comprises a pair of inner primers (FIP, BIP), a pair of outer primers (F3, B3) and a pairof loop primers (LF, LB) with 5' ends marked with fluorescein isothiocyanate and biotin respectively. The application method is characterized in that field rapid detection of the erwinia stewartii pathogenic bacteria can be realized through a loop-mediated isothermal amplification transverse flowing test paper strip detection system. The invention provides a rapid detection technical means for the erwinia stewartii pathogenic bacteria and has strong specificity and high sensitivity.
Description
technical field [0001] The invention relates to a kit and a detection method for rapid detection of bacteria, in particular to a detection method for a loop-mediated isothermal amplification lateral flow test strip for detection of bacterial wilt of corn, which belongs to bacterial wilt of corn detection field. Background technique [0002] Bacterial wilt of corn ( Pantoea stewartii subsp. Stewartii , Pss) is a yellow, immobile, Gram-negative facultative anaerobic bacterium. The host is mainly sweet corn, but also infests dent, flour, durum, and popcorn. All growth stages of corn may be infected by bacterial wilt of corn, and the typical symptoms are dwarfing and wilting. The plant is infected with the disease at the seedling stage, and its growth is severely hindered, resulting in dwarfing and shrinking. The typical symptoms are water-soaked, long gray-green to yellow stripes on the leaves. With the development of the disease, the stripes gradually wither, and the whole ...
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Login to View More IPC IPC(8): C12Q1/689C12Q1/04C12Q1/6844C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2531/119
Inventor 殷宏邹明强薛强何彦平汤小玉王升
Owner CHINA INSPECTION LAB TECH CO LTD