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Human body important parasite antigen chip and method for making same

A parasite antigen and chip technology, applied in the biological field, can solve problems such as low efficiency, slow speed, and lack of specificity, and achieve the effects of easy use, improved reliability, and simple operation

Inactive Publication Date: 2009-01-07
ZHEJIANG ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of specificity in the clinical manifestations of these parasitic diseases, when it is difficult for medical staff to determine what kind of parasitic infection it is, they need to use a single kit to test one by one. Each test can only complete one indicator, which is not only slow, inefficient, and The cost is high, and the amount of samples to be tested must be large

Method used

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  • Human body important parasite antigen chip and method for making same
  • Human body important parasite antigen chip and method for making same

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Microarray detection method for important human parasite antigens

[0038] 8 kinds of important human parasite antigens and 1 internal reference quality control human IgG, a total of 9 protein samples, set 3×3 spotting array, spotting volume 100nl, antigen spot diameter 1.5mm, spot spacing 1.2mm, antigen array The size is 7.5mm×7.5mm, the chip size is 12mm×12mm; 3×3 antigen spot array, as shown figure 2 .

[0039] Cut a certain area of ​​nitrocellulose membrane with a pore size of 0.45 μm according to the number of prepared antigen chips. Clonorchis sinensis adult soluble antigen (Cls), Paragonimus westermani adult soluble antigen (Pw), Schistosoma japonicum egg soluble antigen (Sj), Angiostrongylus cantonensis adult worm antigen (Ac), Dilute Sparganum mansoni soluble antigen (Sm) and Trichinella larvae soluble antigen (Ts) to 0.5 mg / ml, dilute cystic cyst fluid crude antigen (Cyt) to 0.2 mg / ml, and dilute Toxoplasma gondii antigen (Tox) Dilute to 2.0mg / ml, human Ig...

Embodiment 2

[0042] Patient Serum Sample Testing

[0043] Take out one reactor equipped with the antigen chip, put it on the horizontal platform, add eluent (PH7.0, 0.02M TBS, 0.05% sodium azide, 0.05% spit Wen-20, 0.1% Triton-100) 50μl, after fully infiltrated, add 50μl of serum of the patient to be tested, after fully infiltrated, add 50μl of eluent, after fully infiltrated, add 50μl of gold standard solution, after fully infiltrated, add washing Remove 50 μl of the liquid, and after it has completely infiltrated, the results can be read with the naked eye or scanned with a scanner and saved. Each operation step should be carried out continuously, and the entire detection process only takes 3 to 5 minutes. Judgment of results: the internal reference quality control point located in the center shows a small red dot, which means that the test is valid, and if no red dot is displayed, it indicates that the test is invalid; observe the positions of the 8 corresponding parasite antigen spots...

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Abstract

The invention relates to the technical field of biology, in particular to an antigen chip used for detecting human parasites antigens in serum and a preparation method thereof. The invention spots antigen array of parasites which are seriously harmful to human body, Schistosoma, Lung fluke, Clonorchis sinensis, Cysticercosis, Sparganum mansoni, Trichina, Angiostrongylus cantonensis, Toxoplasma gondii, and the like, on a solid phase membrane carrier, and specific gamma immunoglobulin (IgG) of human parasites in serum is detected by a specially made vertical flow chip detect device, and the detecting marker is colloid gold second antibody conjugate or colloid gold protein A conjugate of fresh color. IgG antibodies in serum specially bind with the antigens on the membrane in percolation process layer upon layer, and human IgG which specially binds with the antigens is detected by the colloid gold marker. Detecting of a plurality of parasites antigen indexes in serum sample can be finished within several minutes. The device and the method have the advantages of high throughput, simple operation, convenient use and being suitable for parasitic diseases clinical test and serum-epidemiological investigation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an antigen chip for detecting antibodies to important human parasites in serum and a preparation method thereof. Background technique [0002] Schistosomiasis japonicum is still prevalent in 7 provinces and cities in my country, and the epidemic has rebounded in some areas. According to statistics, the number of people infected with schistosomiasis in my country has reached more than 800,000. With the improvement of people's living standards, social factors such as dietary diversification and fresh dishes have led to significant changes in the prevalence of food-borne parasitic diseases. Some food-borne parasitic diseases that have been basically controlled, such as Clonorchis sinensis Paragonimiasis, paragonimiasis and previously rare foodborne parasitic diseases such as angiostrongylus cantonensis and sparganosis mansoni re-epidemic or small-scale outbreaks occurred. According to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543
Inventor 干小仙王越施晓华
Owner ZHEJIANG ACAD OF MEDICAL SCI
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