Indirect ELISA (Enzyme Linked Immunosorbent Assay) method for detecting anti-roundworm antibody by recombined roundworm ALAg protein antigen

A protein antigen, roundworm technology, applied in the biological field, can solve the problem of missed detection rate and other problems, and achieve the effect of high sensitivity

Inactive Publication Date: 2011-12-14
GUIYANG COLLEGE OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a recombinant roundworm ALAg protein antigen detection anti-roundworm antibody indirect ELISA method, which is used for clinical monitoring of roundworm infection, so as to overcome the missed detection or detection of the existing technology (fecal egg detection method) low rate

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  • Indirect ELISA (Enzyme Linked Immunosorbent Assay) method for detecting anti-roundworm antibody by recombined roundworm ALAg protein antigen
  • Indirect ELISA (Enzyme Linked Immunosorbent Assay) method for detecting anti-roundworm antibody by recombined roundworm ALAg protein antigen
  • Indirect ELISA (Enzyme Linked Immunosorbent Assay) method for detecting anti-roundworm antibody by recombined roundworm ALAg protein antigen

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Embodiment 1

[0035] Example 1. Follow the steps below:

[0036] 1. Preparation of infected infectious eggs

[0037] The fresh feces of children suffering from ascariasis (diagnosed by feces examination of eggs) were collected by repeated sedimentation and placed on filter paper soaked in 2% formalin prepared in advance. Then put the filter paper in a petri dish with wet cotton as the bottom, cover it, and cultivate it in an incubator at 25-28°C, and a large number of infectious eggs can be obtained in 3-4 weeks (during the cultivation process, in order to prevent formaldehyde For the volatilization of forest, a certain amount of 2% formalin solution and normal saline should be added frequently).

[0038] 2. Extraction and RT of total RNA of roundworm adult worms

[0039] Take out the infectious eggs from the refrigerator at -70°C, and extract the total RNA of adults according to the instructions of the RNA extraction kit; use the extracted total RNA as a template, use Oligo dT(18) as a ...

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Abstract

The invention discloses an indirect ELISA (Enzyme Linked Immunosorbent Assay) method for detecting anti-roundworm antibody by a recombined roundworm ALAg protein antigen. In the method, RNA (Ribonucleic Acid) of a roundworm adult body or infective egg serves as a template; an ALAg gene is amplified by RT-PCR (Reverse Transcriptase-Polymerase chain Reaction); an ALAg expression carrier is built, and expression bacteria are transformed for inducible expression; recombined protein is purified; and the indirect ELISA method is built by taking the purified protein as a coated antigen. A stable and specific indirect ELISA method for an anti-roundworm body IgG is built, can be used for the clinical monitoring and diagnosis for roundworm infection and is suitable to basically and clinically widely popularize.

Description

technical field [0001] The invention relates to a prevention and detection technology of roundworm, in particular to the preparation of recombinant roundworm ALAg protein antigen and the indirect ELISA method for detecting anti-roundworm antibody, belonging to the field of biotechnology. Background technique [0002] Roundworm (Ascaris lumbricoides Linnaeus, 1758) is also called ascaris lumbricoides. Ascaris lumbricoides is distributed worldwide and has become one of the public health problems. According to reports, about 1.472 billion people in the world are infected with Ascaris lumbricoides. my country is one of the countries where the infection of Ascaris lumbricoides is more serious. In 1990, the infection rate of Ascaris lumbricoides was 44.59% according to the national survey on the distribution of human parasites. The distribution range is all over the country, especially in rural areas. larger. In 2008, Sun Fenghua reported that a total of 356,629 people were exami...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531C12N15/12C12N15/70C07K14/435
Inventor 何光志田维毅安传伟王平黄高王文佳俞琦奚锦王乾宇
Owner GUIYANG COLLEGE OF TRADITIONAL CHINESE MEDICINE
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