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Schistosome antigen for inducing short-lived antibody response and schistosomiasis diagnostic kit and detection method for detecting antibody response

A technology of short-lived antibodies and schistosomiasis, which is applied in the fields of botany equipment and methods, biochemical equipment and methods, applications, etc., and can solve problems such as inability to diagnose patients

Inactive Publication Date: 2014-03-26
JIANGSU INST OF PARASITIC DISEASES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Soluble egg antigen (SEA) is a mixture composed of many antigenic components. Using SEA as an antibody to detect antigens has certain disadvantages, such as cross-reaction with other parasite antigens, and anti-SEA antibodies in patients remain after they are cured. Can exist for a long time, even a lifetime
The detection of anti-SEA antibodies can only be used for the screening of patients with a history of schistosomiasis infection, not for the diagnosis of patients who are currently infected

Method used

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  • Schistosome antigen for inducing short-lived antibody response and schistosomiasis diagnostic kit and detection method for detecting antibody response
  • Schistosome antigen for inducing short-lived antibody response and schistosomiasis diagnostic kit and detection method for detecting antibody response
  • Schistosome antigen for inducing short-lived antibody response and schistosomiasis diagnostic kit and detection method for detecting antibody response

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Preparation of Schistosoma japonicum adult excreted and secreted antigen

[0077] Rabbits were infected with 1500 cercariae of Schistosoma japonicum. After 42 days, the rabbits were perfused with anticoagulant saline through the portal vein, and the adult worms were collected. Wash the adults repeatedly with sterile PBS to remove surface attachment proteins, place them in cold deionized water, place them in a 37°C incubator for 15 minutes, and take out the adults. The remaining culture was centrifuged at 12000g at 4°C for 30 minutes to remove the precipitate, the supernatant was lyophilized and concentrated, and the protein concentration was determined by BROADFORD method.

Embodiment 2

[0078] Example 2 Preparation of serum samples at different times before and after treatment of schistosomiasis-infected rabbits

[0079] Three healthy young New Zealand white rabbits were taken, and blood was collected through the ear vein before infection with schistosome, and the serum of rabbits without infection was collected. Then each rabbit was infected with 200 cercariae of Schistosoma japonicum through the abdominal skin patch method, and blood was collected through the ear vein at 2 weeks, 4 weeks and 6 weeks after infection, and the serum was separated. At 6 weeks (42 days) after infection, the infected rabbits were treated with praziquantel (350 mg / kg) orally for 2 consecutive days, and then blood was collected every 2 weeks to collect serum until 16 weeks after treatment.

Embodiment 3

[0080] Example 3 Two-dimensional electrophoresis immunoblotting method to identify excreted and secreted antigenic components of adult worms that induce short-lived antibody responses. With the same amount of Schistosoma japonicum adult worms excreted and secreted antigenic protein (1 mg), six 24cm gel strips (IPG, pI 3-10 ) of two-dimensional electrophoresis. After electrophoresis, one of the gel SDS-PAGE gels was stained with Coomassie brilliant blue ( figure 1 a), the other 5 gels were subjected to immunoblotting electrotransfer, and the protein spots separated on each gel were transferred to 5 nitrocellulose membranes of the same size. Then, block with 5% non-fat dry milk in PBS. For the sealed NC membrane, one piece of glue reacted with the serum of healthy rabbits before infection, one piece of glue reacted with the serum of rabbits 6 weeks after infection, one piece of glue reacted with serum of rabbits 4 weeks after treatment, and one piece of glue reacted with serum...

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Abstract

The invention relates to a schistosome antigen for inducing short-lived antibody response and a schistosomiasis diagnostic kit and a detection method for detecting the antibody response, belonging to the technical field of schistosomiasis prevention and treatment and immunological diagnosis. Protein molecules such as glyceraldehyde-3-phosphate dehydrogenase (SjGAPDH) and the like capable of inducing short-lived antibody response in a schistosomiasis infected person through an immunoblotting technology and a mass spectrometry method are identified. A recombinant SjGAPD protein is prepared by adopting a genetic engineering technology, and an enzyme-linked immunosorbent assay detection kit and detection method for detecting anti-SjGAPDH protein specific short-lived antibody response in the schistosomiasis infected person by the recombinant SjGAPD protein are established to diagnose the schistosomiasis infected person and estimate the treatment effect.

Description

technical field [0001] The invention provides an antigen, preparation, diagnostic kit and detection method for inducing antigen-dependent specific short-lived antibody reactions, and relates to the discovery of Schistosoma japonicum triphosphate glyceraldehyde dehydrogenase (Sj GAPDH) by immunoblotting technology and protein mass spectrometry technology ) and other molecules that can induce specific antigen-dependent short-lived antibody responses in patients with Schistosoma japonicum infection, prepared recombinant Sj GAPDH protein, and established a specific short-lived anti-Sj GAPDH protein for the diagnosis and evaluation of schistosomiasis. The reagent kit for antibody reaction belongs to the technical field of immunological diagnosis. Background technique [0002] Schistosomiasis is one of the most important human tropical diseases. There are 76 countries in the world where schistosomiasis is endemic. It is estimated that there are 20 million schistosomiasis patients,...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/63G01N33/573G01N33/543
CPCC12N9/0008C12N15/70C12Y102/01012G01N33/573G01N2333/90203
Inventor 余传信王玠殷旭仁华万全
Owner JIANGSU INST OF PARASITIC DISEASES
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