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
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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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