Cryptosporidium parvum immune colloidal gold detection test paper strip and production method thereof

A technology for detection of Cryptosporidium parvum and test paper, which is applied in the field of parasite detection, can solve the problems of low detection specificity, low sensitivity, and many errors, and achieves the effects of high sensitivity, strong specificity, and simple and fast operation.

Inactive Publication Date: 2011-09-14
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention discloses a cryptosporidium parvum immune colloidal gold detection test strip, which aims to solve the current shortco

Method used

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  • Cryptosporidium parvum immune colloidal gold detection test paper strip and production method thereof
  • Cryptosporidium parvum immune colloidal gold detection test paper strip and production method thereof
  • Cryptosporidium parvum immune colloidal gold detection test paper strip and production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Cloning, expression and protein purification of Cryptosporidium parvum virus capsid protein gene S-dsRNA

[0027] 1. Materials and methods

[0028] (1) Strains and plasmids

[0029] The pET-28a (+) vector was preserved by our laboratory; Escherichia coli DH5a competent and Escherichia coli BL21(DE3) competent were products of Tiangen Biochemical Technology (Beijing) Co., Ltd.; plasmid pMD18-T was provided by Japan TaKaRa Company; Cryptosporidium parvum The cDNA of total RNA is preserved in our laboratory.

[0030] (2) Related molecular biology operations

[0031] Operations such as PCR amplification, plasmid recombination, and transformation were carried out according to the methods described in "Molecular Cloning"; bacterial plasmid DNA extraction, DNA recovery and purification were carried out according to the kit instructions; DNA sequencing was completed by a bioengineering company.

[0032] Gene amplification and cloning

[0033] According to the S-dsRNA gen...

experiment example 1

[0079] Determination and Practice of Colloidal Gold Immunochromatography Test Strip Performance

[0080] 1. Specificity of colloidal gold immunochromatography test paper for cryptosporidium parvum virus capsid protein

[0081] The same method was used to treat the feces containing Cryptosporidium amrenii, Giardia lamblia, Eimeria tenella and Cryptosporidium parvum, and each 50 μL of the filtrate was added dropwise on the test strip for detection. The results showed that the test The strips tested negative for Cryptosporidium amnesia, Giardia lamblia and Eimeria tenella, and positive for Cryptosporidium parvum, indicating good specificity.

[0082] 2. Sensitivity of colloidal gold immunochromatography test paper for cryptosporidium parvum virus capsid protein

[0083] The filtrate was diluted 40 times, and the colloidal gold test strip was detected to be positive, and when the filtrate was diluted more than 80 times, the test strip was detected to be negative. The sensitivity...

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Abstract

The invention provides a cryptosporidium parvum immune colloidal gold detection test paper strip and a production method thereof. The test paper strip comprises a nitrocellulose membrane, a gold marking pad and an absorption pad, wherein the gold marking pad and the absorption pad are arranged at two ends of the nitrocellulose membrane; the upper end of the gold marking pad is a sample pad, and the gold marking pad is coated with a purified CSpV-S protein monoclonal antibody colloidal gold coupling marker, a detection line is coated with a purified monoclonal antibody, a quality control line is coated with a goat anti mouse IgG antibody, and the absorption pad is attached to the side of the quality control line. The research combines the enzyme-immunoassay principle and colloidal gold chromatography to prepare the colloidal gold test paper strip for detecting C.parvum, which is going to be applied to the clinic to improve the prevention and treatment ability of C.parvum, and the invention has the advantages of simple and rapid operation, clear detection results and easy judgment, high specificity, high sensitivity, no need of instruments and apparatuses or just need of simple instruments and the like, and therefore is especially suitable for clinic sample detection use in sites where diseases occur, outpatient departments, places having no experimental conditions and the like.

Description

technical field [0001] The invention provides a test strip for detecting Cryptosporidium parvum immune colloidal gold, which is used for the detection of Cryptosporidium parvum and Cryptosporidium parvum. The invention also discloses a preparation method of the test strip, which belongs to the technical field of parasite detection . Background technique [0002] Cryptosporidium is a ubiquitous gastrointestinal parasitic protozoan that can cause mass diarrhea in humans and livestock. The feces of humans, domestic animals, pets and wild animals containing Cryptosporidium oocysts directly threaten the safety of drinking water, leading to large-scale waterborne outbreaks of Cryptosporidiosis. Molecular epidemiological studies have shown that most human cryptosporidiosis is caused by Cryptosporidium hominis and Cryptosporidium parvum. All Cryptosporidium and Cryptosporidium parvum contain Cryptosporidium parvum virus, which is spherical and about 31 nm in diameter. It is a dsRN...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/532C12N15/30C12N15/70C07K14/44C07K16/20
Inventor 宫鹏涛张西臣刁玉梅李建华张国才杨举张楠李赫邢沈阳
Owner JILIN UNIV
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