Method and kit for detecting tick pathogen nucleic acid based on suspension array

A liquid phase chip, pathogen technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of certain result judgment, insufficient coverage, complicated operation, etc., to achieve low environmental interference and sample volume. Less, better repeatability

Active Publication Date: 2021-09-28
黑龙江省牡丹江林业中心医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some current non-diagnostic detection methods have disadvantages such as low sensitivity, complicated operation, and certain subjectivity in the judgment of results, and almost all of them adopt the detection of a single pathogen, and the coverage is insufficient, which may easily lead to missed detection.

Method used

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  • Method and kit for detecting tick pathogen nucleic acid based on suspension array
  • Method and kit for detecting tick pathogen nucleic acid based on suspension array
  • Method and kit for detecting tick pathogen nucleic acid based on suspension array

Examples

Experimental program
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Effect test

Embodiment 1

[0075] 1. Liquid phase chip preparation

[0076] Take out 40u1 (1×105 pieces) NovaStar magnetic fluorescent microspheres with surface carboxyl modification developed by Wuhan Xinzongke Virus Disease Engineering Technology Co., Ltd., centrifuge at 12000rpm for 2min, discard the supernatant, and add 5u10.1M, pH 4.5 MES to the precipitate solution (2-(N-morpholino)ethanesulfonic acid), and mix well to obtain a coupling system. Dilute the Tag probe solution to 0.1 mM, and add 1 ul to the coupling system. Then add 2.5u1 l0mg / ml EDC (dichloroethane), mix well and place it in the dark for 30min. Add 2.5u1 l0mg / ml EDC again, mix well and place in the dark for 30min. Wash once with 0.2m1 volume percentage of 0.02% Tween-20 and mass volume ratio of 0.1% SDS sodium dodecyl sulfate) solution, and finally resuspend the microspheres in 10u11×TE (pH8.0, the composition of which 1M Tris-HCl, 1mL; 0.5mM EDTA, 0.2mL, plus 100mL of double distilled water) solution to obtain fluorescent micros...

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Abstract

The invention belongs to the technical field of tick detection, and particularly relates to a method and kit for detecting tick pathogen nucleic acid based on a suspension array. The method for detecting tick pathogen based on the suspension array comprises the following steps of S1, designing a PCR primer, a Tag probe sequence and a double-binding probe sequence, wherein the PCR primer is used for amplifying six target gene segments and one reference gene, and the Tag probe is a probe which is modified by AminolinkerC12 at the 5' tail end, is designed aiming at the six target genes and the reference gene and is used for marking the fluorescent microspheres. According to the method and the kit for detecting the tick pathogen nucleic acid based on the suspension array, high-flux multiple detection is performed on six common tick-borne pathogens by adopting a suspension array technology, and the method and the kit have the advantages of high detection speed, high sensitivity, complete pathogen coverage and the like, and can be used as a favorable diagnostic tool for tick detection and evaluation of human and animal infection.

Description

technical field [0001] The invention relates to the technical field of tick detection, in particular to a method and a kit for detecting nucleic acid of tick pathogens based on a liquid phase chip. Background technique [0002] Ticks are the second largest infectious disease vector after mosquitoes. They can transmit a variety of zoonotic diseases and are extremely harmful. Tick-borne pathogens include bacteria, viruses, rickettsia, spirochetes, parasites, etc. There are various types of pathogens, but the current detection methods for tick-borne pathogens are very limited, and there is a lack of relevant commercial detection kits. Some current non-diagnostic detection methods have shortcomings such as low sensitivity, complicated operation, and certain subjectivity in the judgment of results, and almost all of them adopt the detection of a single pathogen, with insufficient coverage and easy to cause missed detection. [0003] MASA liquid chip (Multi-Analyte Suspension Arr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6837C12Q1/6888
CPCC12Q1/6837C12Q1/6888C12Q2600/166C12Q2600/16C12Q2563/107C12Q2563/149C12Q2531/113C12Q2537/143C12Q2545/101C12Q2563/131Y02A50/30
Inventor 边才姚囡囡徐晓微杨喜魁席晓凤叶金玲齐欣红宋聚良郑晓敏郑元春卜晓峰慕义江佳富蒋宝贵
Owner 黑龙江省牡丹江林业中心医院
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