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Kit for detecting Lyme bacteria in tick acarina and detection method thereof

A kit and technology for Lyme disease are applied in the field of kits and detection fields for detecting Lyme disease pathogens in ticks, and can solve the problems of inability to detect mixed infections at the same time, low sensitivity, and single detection of pathogens.

Inactive Publication Date: 2009-12-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

[0006] The present invention aims at detecting the pathogen Borrelia burgdorferi which is the pathogen of Lyme disease in ticks in the prior art when the pathogen is single, the occurrence of mixed infection cannot be detected at the same time, and the sensitivity is low, and the invention provides a direct, comprehensive and highly specific method. and sensitive detection kits for spirochete infection in all currently known samples, and the present invention provides a detection method using this kit at the same time

Method used

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  • Kit for detecting Lyme bacteria in tick acarina and detection method thereof
  • Kit for detecting Lyme bacteria in tick acarina and detection method thereof
  • Kit for detecting Lyme bacteria in tick acarina and detection method thereof

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Embodiment Construction

[0014] 1. Design and synthesis of primers and probes

[0015] Design a pair of primers with a fragment size of 225bp in the conserved region at both ends of the spirochete 5S-23S rRNA gene sequence, and the upstream primers are:

[0016] 23SN2 (5'-ACCATAGACTCTTATTACTTTGACCA-3'),

[0017] The downstream primer is: 5SCB (5'-biotin-GAGAGTAGGTTATTGCCAGGG-3).

[0018] According to the determined variation region of the Lyme disease Borrelia burgdorferi 5S-23S rRNA gene sequence, the design targets ① B. burdorferi sensu stricto, ② B. garinii, ③ B. afzelii and ④ B. burgdorferi Four specific oligonucleotide probes for valaisiana, namely: SS, BG, BA and BV, in addition, a general probe SL for all spirochetes were also designed and synthesized. All oligonucleotide probes are modified with N-(trifluoroacetamidohexyl-cyanoethyl, N, N-diisopropylphosphoramidite[TFA])-C6 amino groups, and the synthesized probes are 0-800pmol / 150ul500mM NaHCO 3 (pH 8.4), to find the optimal concentration ...

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Abstract

The invention relates to a kit for detecting Lyme bacteria in tick acarina and a detection method thereof. The kit for detecting the Lyme bacteria in the tick acarina at least comprises: Biodyne C membrane covalently bonded with universal probes of Lyme bacteria spirochetes and probes for specific oligonucleotide, B. burgdorferi sensu stricto, B. garinii, B. afzelii and B. valaisiana and a pair of primers which are designed on conservative regions at two ends of a gene sequence of spirochete 5S-23S rRNA and have 225 bp fragment that can be amplified, one primer of which is marked with biotin.

Description

technical field [0001] The invention relates to a detection kit and a detection method for detecting a certain pathogen in ticks, specifically an RLB detection method for detecting Lyme disease pathogens in ticks. Background technique [0002] Lyme disease, also known as Lyme soft body disease or Lyme borreliosis, is a tick-borne natural foci of zoonotic spirochetes, which was first diagnosed in Lyme Township, Connecticut, USA. Discovered and named. The pathogen is Borrelia burgdorferi, which can be divided into 10 genotypes. The reported Lyme disease spirochetes in my country include Borrelia burgdorferi sensus stricto, B. garinii and B. garinii Three genotypes of B. afzelii. At present, it has been reported in more than 30 countries and regions in the world that the disease and natural foci exist. The annual incidence is about 300,000, and there is still an increasing trend. It is called "the second AIDS" in the United States (Gratz N. Emerging and resurging vector-borne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/01
CPCY02A50/30
Inventor 殷宏罗建勋牛庆丽关贵全杨吉飞高金亮马米玲刘爱红刘志杰李有全刘军龙任巧云
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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