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Echinococcus multilocularis circulating antigen dot immunogold filtration kit and preparation method

A technology of Echinococcus multilocularis and immunofiltration, which is applied in the field of Echinococcus multilocularis circulating antigen immunofiltration detection kit and its preparation, can solve the problems of inappropriate efficacy assessment and inability to distinguish current infection from past infection, etc. , to achieve the effect of simple and rapid detection, low cost and strong specificity

Inactive Publication Date: 2012-07-25
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since circulating antibodies can still exist for a long time after the treatment of patients, it is impossible to distinguish current infection from past infection, so it is not suitable for efficacy evaluation

Method used

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  • Echinococcus multilocularis circulating antigen dot immunogold filtration kit and preparation method
  • Echinococcus multilocularis circulating antigen dot immunogold filtration kit and preparation method
  • Echinococcus multilocularis circulating antigen dot immunogold filtration kit and preparation method

Examples

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Effect test

preparation example Construction

[0044] The preparation method of the echinococcus multilocularis circulating antigen dot gold immunofiltration kit comprises the following steps:

[0045] 1) Preparation of Echinococcus multilocularis antigens EM2 and EM18

[0046] Using gene cloning technology, PCR amplified DNA encoding Echinococcus multilocularis antigens, and inserted into Escherichia coli to express Echinococcus multilocularis recombinant antigens EM2 and EM18.

[0047] 2) Preparation of monoclonal antibodies against Echinococcus multilocularis recombinant antigens EM2 and EM18

[0048] The recombinant antigen EM2 was mixed with an equal volume of Freund's complete adjuvant to immunize mice, the splenocytes of immunized mice and SP2 / 0 mouse myeloma cells were fused, and the hybridoma cells secreting high-titer antibodies were screened for expanded culture, Freeze. Hybridoma cells were cloned by limiting dilution method. Mice were intraperitoneally injected with positive hybridoma cells. When the abdom...

Embodiment 1

[0079] Coat the antibody against the circulating antigen of Echinococcus multilocularis at the detection point of the nitrocellulose membrane, coat the antibody against the circulating antigen of Echinococcus multilocularis at the detection point, and fix the spotted microporous membrane membrane on the water-absorbing medium Then put it into a carrier medium to assemble a detection plate, one side of the carrier medium has a through hole opposite to the microporous filter membrane, the prepared colloidal gold immunofiltration detection plate is sealed at room temperature and stored for later use. Wherein, the antibody concentration of anti-Echinococcus multilocularis circulating antigen is 1mg / mL, is anti-Echinococcus multilocularis circulating antigen EM2 monoclonal antibody and anti-Echinococcus multilocularis circulating antigen EM18 monoclonal antibody with volume ratio 1: 1 mix; the final concentration of goat anti-mouse IgG polyclonal antibody is 1mg / mL; the sample volum...

Embodiment 2

[0085] Similar to Example 1, the difference is that the anti-Echinococcus multilocularis circulating antigen antibody coated at the detection point of the nitrocellulose membrane is only composed of anti-Echinococcus multilocularis circulating antigen EM2 monoclonal antibody, and does not contain anti-Echinococcus multilocularis Monoclonal antibody to circulating antigen EM18. Result judgment is identical with embodiment 1.

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Abstract

The invention provides an Echinococcus multilocularis circulating antigen dot immunogold filtration kit and a preparation method, relating to an Echinococcus multilocularis circulating antigen detection reagent. The kit is provided with a detection plate, a gold-marked antibody for resisting an Echinococcus multilocularis circulating antigen, and a washing solution; and the detection plate is provided with a carrier medium, a micro-pore filtration membrane, a water absorbing medium, an Echinococcus multilocularis circulating antigen detection point and a contrasting point. The preparation method comprises the following steps of: preparing the Echinococcus multilocularis circulating antigen; preparing the antibody for resisting the Echinococcus multilocularis circulating antigen; preparing colloidal gold; marking the colloidal gold and the antibody for resisting the Echinococcus multilocularis circulating antigen; and preparing the dot immunogold filtration kit. The Echinococcus multilocularis circulating antigen dot immunogold filtration kit can be used for detecting the Echinococcus multilocularis circulating antigen in samples including whole blood, blood serum, blood plasma and the like. When the Echinococcus multilocularis circulating antigen dot immunogold filtration kit is used for detecting, the needed sample amount is extremely small, a special instrument is not needed and a result can be directly judged and read by eyes; and the Echinococcus multilocularis circulating antigen dot immunogold filtration kit has the advantages of simplicity and rapidness in detection, strong specificity, high flexibility, accuracy and reliability, low cost and wide application.

Description

technical field [0001] The invention relates to a detection reagent for Echinococcus multilocularis circulating antigen, in particular to an immunofiltration detection kit for Echinococcus multilocularis circulating antigen by colloidal gold immunofiltration technology (immunofiltration) and a preparation method thereof. Background technique [0002] Alveolar echinococcosis (AE) is a zoonotic parasitic disease caused by the larvae of Echinococcus multilocularia (Em) of the genus Echinococcus, mainly distributed in Inner Mongolia and Xinjiang in my country. The vast pastoral areas such as Tibet, Gansu, Ningxia, Qinghai and western Sichuan are one of the main parasitic diseases currently being controlled in my country. The disease is considered the deadliest helminth infection and one of the most dangerous "zoonotic diseases" worldwide. Humans are infected by ingesting Echinococcus eggs, and then the larvae infiltrate, migrate and grow in the patient's liver in a multivesicula...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531G01N33/532
Inventor 刘凡石松林林丽蓉
Owner XIAMEN UNIV
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