Diagnostic reagent kit for trichinosis by employing dot-immunogold filtration assay

Abstract

The invention discloses a diagnostic reagent kit for trichinosis by employing dot-immunogold filtration assay. The invention has the following advantages: gold labeled SPA is taken as an antibody labeling tag, isopropyl beta-D-thiogalactoside is used for inducing the pMAL-c2X-Ts21 / TB1 recombinant strain, expressing and purifying trichina Ts21 gene-encoded protein (Ts21 antigenic gene, ORF17.20, the access number at GenBank is U88239), after being purified on a nitrocellulose membrane (NCM), the Ts21 recombinant protein coats the detection point T, and human IgG coats the quality control point C, thus constructing Ts21 recombinant protein-dot-immunogold filtration assay (Ts21-DIGFA) and assembling the rapid diagnostic reagent kit for trichina with Ts21 recombinant protein by employing dot-immunogold filtration assay. The kit is simple to operate and has high sensitivity and specificity and good stability and repeatability; the detection results can be observed and judged within 1min immediately after adding samples, the diagnosis results are accurate and clear and misjudgment can not happen; and the kit is suitable for rapid diagnosis and serum epidemiological survey at the scene and is easy to popularize and apply extensively.

Description

technical field [0001] The invention relates to immune detection technology, in particular to a dot immunogold filtration method diagnostic kit for detecting anti-trichinella antibody in serum with trichinella spiralis Ts21 recombinant antigen. Background technique [0002] Trichinellosis is a serious zoonotic disease, which is mainly infected by eating raw or half-cooked pork or other animal meat containing Trichinella larva cysts. This disease not only poses a serious threat to human health, but also The pig industry and the meat import and export trade cause huge economic losses. [0003] At present, the clinical diagnosis of trichinellosis mainly depends on muscle biopsy, but it is often difficult to detect mild and early infections. Even in advanced patients, the positive rate of biopsy is only about 50% due to limited material collection, and it is not easy to be accepted by patients; A lot of research has been done on the serological diagnosis methods of insect disea...

AI Technical Summary

Problems solved by technology

[0005] Dot-immuogold filtration assay (DIGFA) based on colloidal gold labeling technology is a new method established in recent years for the detection of various viruses, bacteria, parasitic diseases and autoimmune diseases. It is an indirect method or a sandwich method (indirect method for antibody detection: specific antigen immobilized on the membrane + corresponding antibody in the specimen + gold-labeled antibody or aureus A (SPA) for color development; sandwich method for antigen detection: fixed on The polyclonal antibody on the membrane + the antigen to be tested in the specimen + the gold-labeled specific monoclonal antibody is developed), the detection speed of the dot immunogold diafiltration method is fast, and the result observation is clear at a glance. It has the advantages of convenient production, low price, easy operation, and no Radioactive contamination and other advantages, but this method has not been used to detect trichinosis

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