Primer for borreia burgdorferi nucleic acid detection, kit, application and detection method

A Borrelia burgdorferi detection method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of unconfirmed genotype pathogenicity correlation

Pending Publication Date: 2020-10-27
杭州美康盛德医学检验实验室有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is currently recognized that B.burgdorferi s.s, B.garinii, B.afzelii, B.bavariensis and B.bissettii are pathogenic, and as B.valaisiana, B.lusitaniae, B.kurtenbachii and B.spielmanii have been isolated from patients one after another It is also considered to be pathogenic, but the pathogenicity of some genotypes and their association with related clinical symptoms have not been confirmed

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  • Primer for borreia burgdorferi nucleic acid detection, kit, application and detection method
  • Primer for borreia burgdorferi nucleic acid detection, kit, application and detection method
  • Primer for borreia burgdorferi nucleic acid detection, kit, application and detection method

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Embodiment example

[0034] Serum and plasma samples from patients with Lyme disease were collected, and the Borrelia burgdorferi fragment gene synthesis plasmid was used as a positive control, and normal human serum was used as a negative control. You can also choose to collect human physiological tissue as a test sample, such as skin or rash exudate.

[0035] (1) extraction of sample nucleic acid; (2) sample addition: add sample nucleic acid, positive control or negative control into PCR tubes equipped with PCR reaction system respectively to obtain corresponding sample reaction tubes, positive reaction tubes or negative reaction tubes, The PCR reaction system contains the aforementioned Borrelia burgdorferi detection primers and probes; (3) PCR reaction: the reaction tube is placed on the PCR instrument, cycle parameters are set, and the PCR reaction is carried out; (4) After the PCR reaction is completed, analyze the results.

[0036] Preferably, in step (3), the conditions of the PCR reaction...

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Abstract

The invention discloses a primer for borreia burgdorferi nucleic acid detection, a kit, application and a detection method. The primer is characterized by comprising a set of specific primers and a probe, the sequences of the set of specific primers are respectively: a primer 1: 5'-TCTTTTCTCTGGTGAGGGAGCT-3'; a primer 2: 5'-CCTTCCTGTTGAACACCCTCT-3'; and the sequence of the probe is: 5'-FAM-AAACTGCTCAGGCTGCACCGGTTC-BHQ1-3'. Multiple subtypes of borreia burgdorferi can be detected by using the specific set of primers and the probe, and the primer has the advantages of simplicity, convenience, rapidity, accuracy and efficiency.

Description

technical field [0001] The invention relates to a primer, a kit, an application and a detection method thereof for Borrelia burgdorferi nucleic acid detection, the sample is sealed and cross-contamination is avoided; the kit has strong anti-interference effect, strong specificity and specificity, and avoids false positives Results: The protozoa rate of the minimum detection limit of the present invention can reach 1%, and the human housekeeping gene is used as the internal standard quality control product in the amplification process, so as to avoid additional addition of nucleic acid extraction and reduce the risk of human interference. Background technique [0002] Lyme disease is a spirochete infectious disease mediated by ticks, which is a natural foci disease caused by Borrelia burgdorferi. In 1985, my country first discovered the disease case in the forest area of ​​Heilongjiang Province, and the main clinical manifestation of the disease was nervous system damage. Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11C12R1/01
CPCC12Q1/689C12Q1/686C12Q2600/166C12Q2561/113C12Q2563/107
Inventor 金永楠肖兰刘阳
Owner 杭州美康盛德医学检验实验室有限公司
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