Universal primer for detecting blood protozoan Colpodella in body of south China tiger or tick

A universal primer, the technology of South China tiger, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of false positives and so on

Inactive Publication Date: 2020-04-03
LONGYAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, using nested PCR technology to detect the blood protozoa Colpodella in the blood of South China tigers has the disadvantage of false positives

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0037] 1. Sample source

[0038] The blood samples and tick samples were all from the Meihuashan South China Tiger Breeding Base. Blood samples were stored at low temperature in EDTA anticoagulant tubes, and tick samples were stored at low temperature in 50 mL centrifuge tubes.

[0039] 2. Blood DNA Extraction

[0040] (1) Pipette 200 μL of blood sample into a 1.5 mL sterilized centrifuge tube;

[0041] (2) Add 25 μL OB Protease Solution and 200 μL BL Buffer solution, vortex fully

[0042] 15 seconds evenly;

[0043] (3) 65°C water bath for 10 minutes, take out and mix briefly every 5 minutes;

[0044] (4) Add 260 μL of absolute ethanol, thoroughly vortex and mix for 20 seconds, and briefly centrifuge to collect all samples;

[0045] (5) Put the adsorption column in the collection tube, transfer all the liquid into the adsorption column, centrifuge at 12000rpm for 1 minute, discard the filtrate and collection tube, and put the adsorption column into a new collection tube;...

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PUM

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Abstract

The invention discloses a universal primer for detecting a blood protozoan Colpodella in the body of a south China tiger or a tick. The universal primer comprises two groups of primers: Colp-F1 and Colp-R1, and Colp-F2 and Colp-R2. The designed primer provided by the invention has the advantages that the positive proportion and the accuracy of fast screening the blood protozoan Colpodella in a host and an intermediate host can be improved; the defect of false positive rate is avoided; unnecessary sequencing time and money waste can be reduced; and great help is provided for fast screening theblood protozoan Colpodella in the host and the intermediate host.

Description

technical field [0001] The invention belongs to the technical field of parasite molecular detection, and in particular relates to a general primer for detecting blood protozoa Colpodella in South China tigers or ticks. Background technique [0002] Blood protozoa currently detected Colpodella There are few universal primers used, so far there are only two sets of universal primers (Piro1-S and Piro3-AS and PIRO-B and PIRO-A), which are not listed in the international SCI literature. Based on the fact that the South China tiger is a unique subspecies of tiger in my country, it is an irreplaceable existence in nature and has extremely high research value, so it is developed for detection Colpodella It is imperative to use the nested PCR technology to detect South China tiger blood or to prevent and control infectious and highly pathogenic ticks carrying this pathogen. It is also one of the ways to prevent the endangered species of South China tiger. The data basis for the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6893C12Q1/6848C12Q1/04C12N15/11
CPCC12Q1/6893C12Q1/6848C12Q2549/119C12Q2565/125
Inventor 邱泓铨黄翠琴孙晓双李世贤
Owner LONGYAN UNIV
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