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Reagent and kit for detecting plasmodium and distinguishing four plasmodium species through isothermal amplification and application of reagent and kit

An isothermal amplification, Plasmodium technology, applied in the direction of resistance to vector-borne diseases, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of high cost and inconvenient detection, and achieve high sensitivity and false negative. And the effect of low false positive and good repeatability

Pending Publication Date: 2021-12-07
中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of this application is to provide a kind of isothermal amplification detection of Plasmodium and the reagent, kit and application thereof which can distinguish four Plasmodium species. High, the problem of inconvenient detection d

Method used

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  • Reagent and kit for detecting plasmodium and distinguishing four plasmodium species through isothermal amplification and application of reagent and kit
  • Reagent and kit for detecting plasmodium and distinguishing four plasmodium species through isothermal amplification and application of reagent and kit
  • Reagent and kit for detecting plasmodium and distinguishing four plasmodium species through isothermal amplification and application of reagent and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] An isothermal amplification reagent for detecting Plasmodium genus and distinguishing four Plasmodium species

[0103] The reagents include the loop-mediated isothermal nucleic acid amplification primer set and the visualization constant temperature amplification reagent for detecting Plasmodium genus and distinguishing four Plasmodium species, wherein the loop-mediated isothermal nucleic acid amplification primer set and the visualization constant temperature amplification reagent are Specifications are shown in Table 4 above.

[0104] Table 4

[0105]

Embodiment 2

[0107] A kit for detecting Plasmodium genus and distinguishing four Plasmodium species through isothermal amplification

[0108] The kit includes the reagents provided in Example 1, nucleic acid extraction consumables, positive quality controls and negative quality controls;

[0109] Among them, nucleic acid extraction consumables include nucleic acid extraction solution and devices for nucleic acid extraction. The nucleic acid extraction solution is selected from processing solution, lysate, cleaning solution, and eluent. The specific addition amount and selection are shown in Table 2. The device is selected from Liquid container, nucleic acid lysing container, nucleic acid concentrator; and, in the liquid taker 5ml, 1ml screw piston syringe, the nucleic acid lysing container is selected from the lysis tube, and the nucleic acid concentrator is a self-assembly consumable, which is made by punching The silica gel membrane is cut into a regular circular structure, and loaded in...

Embodiment 3

[0112] An isothermal amplification method for detecting Plasmodium genus and distinguishing four Plasmodium species

[0113] The isothermal amplification provided in Example 2 is used to detect the genus Plasmodium and the kit capable of distinguishing four kinds of Plasmodium species to detect the genus Plasmodium and to distinguish four kinds of Plasmodium species. The detection method comprises the following steps:

[0114] S11. Extraction method of sample nucleic acid

[0115] Samples to be tested: 20 copies of whole blood from patients with Plasmodium vivax, whole blood from patients with Plasmodium falciparum, whole blood from patients with Plasmodium malaria, and whole blood from patients with Plasmodium ovale;

[0116] Sample pretreatment: Pipette 200μL-1000μL of the sample to be tested into the treatment tube (containing the treatment solution) with a 1ml pipette, mix well and leave it at room temperature for 5 minutes to perform the first lysis;

[0117] Sample lysi...

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Abstract

The invention relates to the technical field of molecular biological detection, and provides a reagent and kit for detecting plasmodium and distinguishing four plasmodium species through isothermal amplification and application of the reagent and the kit, wherein the reagent for detecting the plasmodium and distinguishing the four plasmodium species through isothermal amplification comprises loop-mediated isothermal nucleic acid amplification primer groups for detecting the plasmodium and distinguishing the four plasmodium species according to SEQ ID No.1-SEQ ID No.30; therefore, the sensitivity for detecting the plasmodium and distinguishing the four plasmodium species is high; the repeatability is good; the false negative and false positive are low; the detection accuracy is high; and result judgment can be directly carried out by observing colour changes with naked eyes.

Description

technical field [0001] The application belongs to the technical field of molecular biology detection, and in particular relates to a reagent, a kit and an application thereof for isothermal amplification detection of Plasmodium and the ability to distinguish four Plasmodium species. Background technique [0002] Malaria is a vector-borne infectious disease caused by Plasmodium parasites entering the blood through the bite of Anopheles mosquitoes. There are four main types of Plasmodium that parasitize humans, namely Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae and Plasmodium ovale. Another zoonotic species, Plasmodium knowlesi, is a relatively rare cause of human malaria. According to WHO's 2020 malaria report statistics: there are currently 87 malaria countries in the world, with an estimated 229 million cases and 409,000 deaths in 2019. About 95% of global malaria deaths occurred in 31 countries, Nigeria (23%) , the Democratic Republic of the Congo (11%),...

Claims

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Application Information

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IPC IPC(8): C12Q1/6893C12Q1/6844C12N15/11
CPCC12Q1/6893C12Q1/6844C12Q2531/119C12Q2563/173Y02A50/30
Inventor 陈家旭陈木新胡坤敏张岩艾琳陈燕旌黄达娜陈军虎边素莹冯欣宇宋鹏胡薇苏川张仁利周晓农
Owner 中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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