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Plasmodium antibody colloidal gold detection kit and preparation method thereof

A detection kit, colloidal gold technology, applied in measurement devices, resistance to vector-borne diseases, instruments, etc., can solve problems such as low work efficiency, reduced reading accuracy, increased complexity, etc., and achieve strong specificity.

Active Publication Date: 2014-07-16
SHANGHAI CHEMTRON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses an ordinary biological microscope for manually shifting and focusing, and the microscope inspectors often suffer from eye fatigue, resulting in reduced reading accuracy and low work efficiency.
The development of immunochromatography technology using nano-colloidal gold as a marker is very mature, but the monoclonal antibody used in the existing malaria detection kit can only detect one antigen, or simply combine several antibodies to the detection area, which increases the production complexity

Method used

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  • Plasmodium antibody colloidal gold detection kit and preparation method thereof
  • Plasmodium antibody colloidal gold detection kit and preparation method thereof
  • Plasmodium antibody colloidal gold detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] The preparation of embodiment 1 kit

[0058] 1. Materials

[0059] 1.1 30 Balb / c mice, 6-8 weeks old, raised by the company itself;

[0060] 1.2 Reagent source: goat anti-mouse IgG polyclonal antibody, HRP-II-pLDH recombinant protein: purchased from MAXMED LABORATORIES INC., anti-HRP-II monoclonal antibody and anti-pLDH monoclonal antibody ( It can be prepared with reference to existing literature reports, such as Hao Wenbo, Li Ming, etc. Preparation and identification of anti-Plasmodium falciparum recombinant HRP-II monoclonal antibody [J]. Chinese Journal of Parasitic Disease Control, 2000, 13(3): 165- 168; Wang Junyun, Bao Yifang, Yang Yuetao, Tang Linhua, Preparation of specific monoclonal antibody against lactate dehydrogenase of Plasmodium falciparum. Chinese Journal of Parasitology and Parasitic Diseases, 2005, 4, etc.)

[0061] Nitrocellulose film, glass fiber paper: purchased from Germany Sartorius (SARTORIUS)

[0062] 2 methods

[0063] 2.1 Preparation of ...

Embodiment 2

[0127] Example 2 Sensitivity Experiment of Plasmodium Antibody Colloidal Gold Colloidal Gold Detection Kit

[0128] Detection method:

[0129] 1. Take the Plasmodium antibody colloidal gold detection kit prepared in Example 1, and place the kit on a horizontal platform.

[0130] 2. Use the sample pipette to draw the sample, then drop 2 drops (about 50-80 μl) of the sample into the sample hole of the kit, and then add 2 drops of normal saline. Use a different pipette for each different sample.

[0131] 3. Observation results: Interpret the results 20 minutes after adding the sample.

[0132] Test sample:

[0133] Prepare quality control reference substances with HRP-II concentrations of 10ng / ml, 15ng / ml, 20ng / ml, and 25ng / ml as samples.

[0134] Configure quality control reference substances with pLDH concentrations of 5ng / ml, 10ng / ml, 15ng / ml, and 20ng / ml as samples.

[0135] Test results:

[0136] HRP-II concentration (ng / ml)

[0137] pLDH concentrati...

Embodiment 4

[0139] Example 4 Specificity Experiment of Plasmodium Antibody Colloidal Gold Detection Kit

[0140] Get the detection kit that example 1 makes, according to the following table add sample detection respectively, detection result is as follows:

[0141] Unit: ng / ml

[0142] sample

[0143] It can be seen that the specificity of the kit is good.

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Abstract

The invention relates to a plasmodium antibody colloidal gold detection kit and a preparation method thereof. The plasmodium antibody colloidal gold detection kit comprises reagent strips, namely a piece of sample filter paper, an immune colloidal gold paper sheet, an immune nitrocellulose membrane and a piece of water absorption paper sequentially from a sample injection end, wherein an HRPII-pLDH bispecific monoclonal antibody marked by colloidal gold is wrapped on the immune colloidal gold paper sheet; an anti-HRPII specific monoclonal antibody and an anti-pLDH specific monoclonal antibody are sprayed in a detection region (T) of the nitrocellulose membrane; and goat-anti-mouse immune globulin (IgG) is sprayed in a quality control region (C) of the nitrocellulose membrane. The plasmodium antibody colloidal gold detection kit can detect Malaria sensitively, specifically and quickly.

Description

technical field [0001] The invention relates to a detection kit, in particular to a malaria parasite antibody colloidal gold detection kit and its preparation. Background technique [0002] Malaria (Malaria) is caused by the transmission of Plasmodium by the bite of a mosquito in the tropics and subtropics. Different Plasmodium causes Plasmodium vivax, Plasmodium malaria, Plasmodium falciparum and Plasmodium ovale respectively. The main manifestations of this disease are periodic attacks, chills, fever, and sweating all over the body. After repeated attacks for a long time, anemia and splenomegaly can be caused. Malaria is more common in summer and autumn, and can occur throughout the year in tropical and subtropical regions, and it is easy to spread. Malaria is still one of the most serious parasitic diseases threatening and endangering people's health in Asia and Africa. At present, there are still 120 million malaria patients in the world, and about 300 million carriers...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/558
CPCY02A50/30
Inventor 张国华
Owner SHANGHAI CHEMTRON BIOTECH
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