Kit for detecting circulating antigen indirect hemagglutination of schistosomiasis and manufacturing method thereof

A technology of schistosomiasis and kits, applied in the field of bioengineering, can solve problems such as inability to distinguish past infection from current infection, low sensitivity of circulating antigens, and inability to conduct curative effect assessment, achieving considerable economic and social benefits, easy promotion, high The effect of practical value

Inactive Publication Date: 2010-11-24
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide an indirect hemagglutination kit for detecting circulating antigens of schistosomiasis and its manufacturing method. The indirect hemagglutination kit for detecting circulating antigens of schistosomes and its manufacturing method should solve the problem of detecting circulating antibodies of schistosomes Inability to distinguish past infection from current infection, inability to evaluate efficacy, and technical problems of low sensitivity and low specificity for detection of circulating antigens

Method used

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  • Kit for detecting circulating antigen indirect hemagglutination of schistosomiasis and manufacturing method thereof
  • Kit for detecting circulating antigen indirect hemagglutination of schistosomiasis and manufacturing method thereof
  • Kit for detecting circulating antigen indirect hemagglutination of schistosomiasis and manufacturing method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Anti-Schistosome japonicum egg antigen (SEA) specific IgY preparation and purification

[0027] 1. Preparation of soluble antigen of Schistosoma japonicum: 7 New Zealand rabbits each infected with 1500 cercariae of Schistosoma japonicum (from China Center for Disease Control and Prevention, Parasitic Disease Prevention and Control Institute) were dissected according to the conventional method after 42 days, and separated and collected from the liver eggs and freeze-dried. Take the dry eggs and weigh them, dissolve them in 0.9% normal saline at a ratio of 1%, place them in a cold soak at 4°C for 4 days, and shake them twice a day for 2 minutes each time. After 4 days, ultrasonic pulverization was performed in an ice bath, 3 minutes each time, with a 3-minute interval, 3 times in total. Then cold soak for 2 days as above, centrifuge at 10,000 rpm at 4°C for 30 minutes, take the supernatant, collect and aliquot, measure the concentration of antigenic protein...

Embodiment 2

[0034] Embodiment 2: The present invention provides a kit for detecting schistosome circulating antigen indirect hemagglutination and its manufacturing method, including freeze-dried anti-SEA-IgY sensitized red blood cells, sensitized red blood cell diluent, specimen diluent, positive control substance, negative As for the control substance, the positive control substance is a standard dilution of Schistosoma japonicum soluble antigen, and the negative control substance is human normal serum.

[0035] Specifically, the sensitized erythrocyte diluent uses double distilled water (DDH 2 O), described sample diluent is 0.9% physiological saline.

[0036] Preparation of positive control standard product: Select 2.5GK healthy rabbits, infect each rabbit with 1500-2000 cercariae according to the conventional method, separate and collect eggs from the liver after 8 weeks and freeze-dry them. Take the dry eggs and weigh them, dissolve them in 0.9% normal saline at a ratio of 1%, place...

Embodiment 3

[0043] Example 3: Detection of Schistosoma Circulating Antigens Indirect Hemagglutination Kit Use:

[0044] 1. Operation steps:

[0045] (1) Prepare sensitized erythrocyte suspension: take freeze-dried sensitized erythrocytes, add 1ml of diluent, and mix well.

[0046] (2) Negative and positive controls should be set up for each test. Positive and negative control sera are lyophilized products, diluted with 100 μl distilled water in each tube before use, and used after fully dissolved.

[0047] (3) Add 100 μl of sample diluent to the first well of the first row of the hemagglutination plate, and add sample diluent to the second to fourth wells. Add 30μl of the serum to be tested in the first well, mix well and suck out 30ul into the second well, mix well in the second well and then serially dilute to the fourth well, discard the excess 30μl after mixing in the fourth well, Serum dilutions from the second to fourth wells were 1:10, 1:20 and 1:40, respectively. Then add 1 dr...

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Abstract

The invention provides a kit for detecting circulating antigen indirect hemagglutination of schistosomiasis and a manufacturing method thereof. The kit is composed of freeze-drying anti-SEA-IgY sensitized erythrocyte, sensitized erythrocyte diluent, sample diluents, positive comparison products and negative comparison products. The method for manufacturing the kit for detecting circulating antigen indirect hemagglutination of schistosomiasis comprises the steps of preparing a schistosoma japonica soluble antigen, performing antigen immunization and collecting eggs, extracting and purifying specific anti-SEA-IgY, preparing hydroformylated red cells, preparing tanned red cells, preparing a specific anti-SEA-IgY antibody sensitized erythrocyte, and preparing freeze-drying red cells. The kit in the invention has the advantages of simple and convenient operation, rapidness, high sensibility, strong specificity and good repeatability.

Description

Technical field: [0001] The invention belongs to the field of bioengineering, and in particular relates to a kit, in particular to an indirect hemagglutination kit for detecting schistosome circulating antigens and a manufacturing method thereof. Background technique: [0002] 1. The importance of immunodiagnosis in the prevention and treatment of schistosomiasis japonicum [0003] Schistosomiasis is a zoonotic parasitic disease widely prevalent in tropical and subtropical regions. About 200 million people in 76 countries and regions around the world are infected, and 600 million people are threatened by infection. my country is an endemic area of ​​schistosomiasis japonicum, which seriously affected people's normal production and life in the early days of the founding of the People's Republic of China. After more than 50 years of prevention and control, remarkable achievements have been made. By 2003, the total number of schistosomiasis patients in China was 843,007, which...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/86
Inventor 陈家旭蔡玉春郭俭
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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