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.
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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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