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Immune chromatographic test paper for detecting plasmodium and preparing method

An immunochromatographic test paper and Plasmodium technology, which can be applied in the direction of measuring devices, analysis materials, and resistance to vector-borne diseases, etc., can solve the problems of impossible large-scale application and high detection cost, and achieve easy industrial production, simple detection methods, The effect of high sensitivity and specificity

Inactive Publication Date: 2007-06-13
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fluorescent quantitative PCR technology cleverly utilizes the advantages of high-efficiency DNA amplification of PCR technology, high specificity of probe technology, high sensitivity of spectral technology and quantitative analysis, and overcomes some shortcomings of conventional PCR qualitative detection, thus Greatly improved the sensitivity and specificity of detection, but due to factors such as the need for special equipment and high detection costs, it is impossible to apply it on a large scale at the grassroots level and on-site

Method used

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  • Immune chromatographic test paper for detecting plasmodium and preparing method

Examples

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Effect test

Embodiment 1

[0044] Embodiment 1, the preparation of the immunochromatography test paper that detects Plasmodium

[0045] Take the immunochromatographic test paper detecting p.f as an example to illustrate the preparation method of the immunochromatographic test paper detecting Plasmodium of the present invention, and the specific process comprises the following steps:

[0046] 1. Preparation of Plasmodium-specific antibodies

[0047] In this example, recombinant Plasmodium falciparum lactate dehydrogenase (rLDHpf) was used as an antigen to prepare Plasmodium-specific antibodies. The specific method is as follows:

[0048] 1) Preparation of Plasmodium antigens

[0049] ① Preparation of recombinant p.f lactate dehydrogenase (LDHpf) antigen

[0050] A pair of primers were designed according to the LDHpf gene sequence of Honduras 1 strain (GenBank accession number: M93720), and the primer sequences were as follows:

[0051] P1 (upstream primer): 5'-CCTCGAATTCATGGCACCAAAAGCAAAAATCGT-3'

[...

Embodiment 2

[0080] Embodiment 2, the detection of Plasmodium and the cross test with other related biological LDH isozymes

[0081] 1. Detection of Plasmodium

[0082] The p.f worm blood was lysed and diluted with PBS solution (pH 7.4) containing 0.2% saponin, so that all the LDH in the erythrocytes were released for sample application. Detect with the immunochromatographic test paper of coated malaria parasite specific antibody (rabbit anti-rLDHpf specific antibody) and quality control antibody goat anti-rabbit IgG prepared in Example 1, the results are all positive (the detection line and the quality control line place are all positive). A precipitation line appears), which is consistent with the expected result.

[0083] 2. Cross-test with other related biological LDH isozymes

[0084] Treat normal human erythrocytes, Toxoplasma gondii, Cryptosporidium, Trichomonas and Escherichia coli BL21 in the same way as step 1. Similarly, the immunochromatographic test paper coated with the Pl...

Embodiment 3

[0086] Embodiment 3, the laboratory examination of the immunochromatographic test paper that detects Plasmodium

[0087] 1. Sample preparation

[0088] The p.f worm blood was lysed and diluted with PBS solution (pH7.4) containing 0.2% saponin, and the worm density was 10, 50, 100 / ul respectively. And Escherichia coli BL21 are also processed in the same way to obtain detection solutions with different analyte contents for future use. Soak the filter paper in the test solution, completely soak it and then dry it to get the filter paper blood sample. The filter paper blood sample is made into a filter paper blood spot with a diameter of 5mm by a punching machine, and the filter paper blood spot is placed in a small hole of an ordinary microplate plate, and a PBS solution containing 0.2% saponin is added, and the blood sample is eluted on a micro shaker. After fully hemolyzed, remove the filter paper for sample application.

[0089] 2. Detection method

[0090] Add 3 drops (ab...

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Abstract

The invention discloses an immunization chromatography test paper for testing plasmodium and the manufacture method. The paper includes sample tray, gold mark tray, fiber cellulose membrane and sopping tray. The fiber cellulose membrane has a detection line and a quality control line. The detection line contains plasmodium specificity antibody. The invention has the advantages of convenience, sensitivity, specificity and rapidity.

Description

technical field [0001] The invention relates to a test paper for detecting malaria parasites and a preparation method thereof, in particular to an immunochromatographic test paper for detecting malaria parasites and a preparation method thereof. Background technique [0002] Malaria is a vector-borne infectious disease that seriously endangers human health, and its pathogen is Plasmodium. There are four main types of Plasmodium that parasitize the human body: Plasmodium vivax (P.v), Plasmodium falciparum (P.f), Plasmodium malariae and Plasmodium ovale. In my country and other countries in the world, P.v and P.f are the most common, and malaria and malaria ovale are extremely rare. [0003] Malaria is one of the five major parasitic diseases in the world, with high morbidity and mortality. Plasmodium sporozoites enter the human body through the bite of malaria mosquitoes, develop and multiply in the liver, produce a large number of merozoites, release them into the blood, a...

Claims

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Application Information

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IPC IPC(8): G01N33/544
CPCY02A50/30
Inventor 端青邱少富朱虹檀华宋立华何君
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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