Method for detecting foot and mouth disease (FMD) non-structural protein 3AB antibody by using dot immuno-gold filtration assay (DIGFA)

A technology of non-structural protein and spot immunity, which is applied in the field of detection of foot-and-mouth disease non-structural protein 3AB antibody by spot immunogold filtration method, and achieves the effects of good specificity, bright color and easy judgment.

Inactive Publication Date: 2010-09-01
INSPECTION & QUARANTINE TECH CENT OF XIAMEN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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Problems solved by technology

Most developing countries, including China, mainly rely on vaccination to prevent FMD, and FMD antibodies can also be detected in animal serum after immunization, and the detection of FMD antibodies by traditional FMD virus antigen coating cannot be distinguished. Animals infected with wild virus or the purpose of immunizing animals with inactivated vaccines

Method used

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  • Method for detecting foot and mouth disease (FMD) non-structural protein 3AB antibody by using dot immuno-gold filtration assay (DIGFA)
  • Method for detecting foot and mouth disease (FMD) non-structural protein 3AB antibody by using dot immuno-gold filtration assay (DIGFA)
  • Method for detecting foot and mouth disease (FMD) non-structural protein 3AB antibody by using dot immuno-gold filtration assay (DIGFA)

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Embodiment Construction

[0030] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0031] 1. In this example, FMD 3AB non-structural protein was first synthesized

[0032] The antigen used for expression primers and expression recombinant plasmid construction is prokaryotic expression of foot-and-mouth disease 3AB protein. The synthesis of FMD non-structural protein 3AB gene was searched and analyzed by GenBank, and the 3AB gene was selected with reference to the published Asian strain (AY687333). According to the degeneracy of codons Based on the codon preference principle of Escherichia coli and Escherichia coli, some rare codons in the 3AB sequence were modified without affecting the amino acid sequence, and divided into 9 long primers, and 12 complementary sequences were designed between each long primer. Two 3AB genes were obtained by overlapping PCR method, and the nucleotide sequences of the genes were:

[0033] SEQ ID NO.1:

[003...

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Abstract

The invention discloses a method for detecting a foot and mouth disease (FMD) non-structural protein 3AB antibody by using dot immuno-gold filtration assay (DIGFA). The method specifically comprises the following steps of: a) synthesizing a foot and mouth disease(FMD) non-structural protein 3AB; b) preparing SPA colloidal gold; c) manufacturing a reaction kit; and d) applying the reaction kit as a carrier, placing antigen sites of the 3AB non-structural protein synthesized in the step a) on an NC membrane, sealing the NC membrane and adding a sample to be detected, and after washing the NC membrane, using the SPA prepared in the step b) as the colloidal gold to mark the protein for detecting the FMD non-structural protein 3AB antibody. The method has the characteristics of simple, convenient and fast operation, no need of special instrument, bright spot color, strong specificity, high sensitivity, easy judgment of results and preservable detected samples.

Description

technical field [0001] The invention relates to a detection method for detecting the foot-and-mouth disease non-structural protein 3AB antibody by using the dot immunogold filtration method (DIGFA). Background technique [0002] Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious infectious disease caused by foot-and-mouth disease virus infection and cloven-hoofed animals. The disease is characterized by blistering and ulceration of the oral mucosa, tongue, lips, nose, hooves, and breast skin. The World Organization for Animal Health (OIE) listed the disease as the first class A zoonosis. Due to the high incidence of the disease, rapid spread and wide range of epidemics, once it breaks out, it is difficult to control and eliminate, seriously affecting animal husbandry production and international trade, and causing huge economic losses. The disease has a long history of epidemics and is widely distributed all over the world. In most developing countries in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531C12N15/42C12N15/70C07K14/09
Inventor 孔繁德林祥梅徐淑菲朱文钏韩雪清吴绍强陈信忠龚艳清梅琳
Owner INSPECTION & QUARANTINE TECH CENT OF XIAMEN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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