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LAMP detection kit and detection primers for pathogenic bacteria of bacterial mumps of pelodiscus sinensis

A detection kit and bacteriostatic technology, applied in the field of LAMP detection kit and detection primers of Chinese soft-shelled turtle bacterial mumps pathogen, can solve the problems that have not been reported, and achieve easy popularization and application, high detection sensitivity and short detection time Effect

Active Publication Date: 2018-12-07
ZHEJIANG INST OF FRESH WATER FISHERIES
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Problems solved by technology

At present, this technique has not been reported in the diagnosis of Chinese soft-shelled turtle bacterial mumps

Method used

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  • LAMP detection kit and detection primers for pathogenic bacteria of bacterial mumps of pelodiscus sinensis
  • LAMP detection kit and detection primers for pathogenic bacteria of bacterial mumps of pelodiscus sinensis
  • LAMP detection kit and detection primers for pathogenic bacteria of bacterial mumps of pelodiscus sinensis

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Embodiment

[0051] 1. Materials

[0052] The Chinese soft-shelled turtle infected with pathogenic Bacillus cereus was collected from a farm in Xiaoshan District, Hangzhou City, Zhejiang Province; the primers used were synthesized by Shanghai Sangon Biotechnology Company; the large fragment of Bst DNA polymerase was purchased from New England BIPolabs Company; dNTP was purchased from promega Company; calcein, manganese chloride, betaine, magnesium sulfate, etc. were purchased from sigma company.

[0053] 2. Extraction of DNA from soft-shelled soft-shelled turtle samples infected with germs:

[0054] (1) Collect 500ml of Chinese soft-shelled turtle breeding pond water with a sterile sampling bag, filter it with a 0.45μm filter membrane, grind the membrane into pieces, add 100-200μL of sterile distilled water, boil in boiling water for 10min, centrifuge at 5000rpm for 1min, and take the supernatant for later use ;

[0055] (2) Take 25-50 mg of Chinese soft-shelled turtle liver, spleen or l...

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Abstract

The invention discloses a LAMP detection kit and detection primers for pathogenic bacteria of bacterial mumps of pelodiscus sinensis; the kit includes specific primers, a reaction buffer liquid, Bst DNA polymerase and a fluorescent dye. The specific primers are composed of external primers, internal primers and ring primers. The kit has the advantages of strong specificity, high sensitivity and convenient operation, can make up for the current situation that mumps occurs frequently in the culture process of the pelodiscus sinensis but the pathogenic bacteria cannot be confirmed, and is suitable for screening and prevention of bacterial mumps of the pelodiscus sinensis.

Description

technical field [0001] The invention relates to the technical field of soft-shelled turtle disease detection, in particular to a LAMP detection kit and detection primers for soft-shelled soft-shelled turtle bacterial mumps pathogenic bacteria. Background technique [0002] Chinese soft-shelled turtle is a kind of aquatic animal with rare and high economic value. In terms of animal classification, soft-shelled turtles belong to the phylum Chordate, the subphylum Chordate, the class Reptilia, the subclass Apore, the order of Turtles, the suborder of Trionychidae, and the family Trionychidae. There are 7 genera and 23 species of soft-shelled turtles in the family of turtles. There are only 2 genera and 3 species in my country. Among them, there are two species of soft-shelled turtles, namely Chinese soft-shelled turtle (Pelodiscus sinensis) and Shanrui soft-shelled turtle (Palea steindachneri). Turtles are the most common. In the 1960s, Japan began to cultivate Chinese soft-sh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12N15/11C12R1/085
CPCC12Q1/6844C12Q1/689C12Q2531/119C12Q2563/107
Inventor 林锋刘莉曹铮陈建明卢淑娟孙丽慧
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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