Cryptosporidium and Giardia detection and quality control kit
A technology for Giardia and Cryptosporidium, which is applied in the field of Cryptosporidium and Giardia detection and quality control kits, can solve the problems of short shelf life and high cost, and achieve long shelf life, low cost, and control of detection costs. Effect
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Embodiment 1
[0016] The Cryptosporidium and Giardia-specific surface proteins were retrieved from the NCBI database, respectively, and the amino acid sequences of the obtained Cryptosporidium and Giardia-specific surface proteins were compared by blastp software for homology. Sexual comparisons with higher similarity were considered as experimental objects. We selected three proteins with higher homology from the surface proteins of Cryptosporidium, named Cowp (its amino acid sequence is shown in SEQ ID.NO.1), Cp41 (its amino acid sequence is SEQ ID.NO. .3) and Cdg6 (its amino acid sequence is shown in SEQ ID.NO.2), two proteins with higher homology were selected from the Giardia surface proteins, and their names were VSP (its amino acid sequence was SEQ ID. NO. 4), G3 (its amino acid sequence is shown in SEQ ID. NO. 5).
[0017] SEQ ID. NO. 1: FTFSGKQCVQSDTAPPNPECPPGTVLENGTCKLIQQVDTICPPGFVEEGNRCVQYLPANKICPPGFNLSGQQCMAPESAELESTCPPNTILENGKCKVIKNVDMVCPPGYTDSGDECVLYVAPAKECPPNFTLQG
[0018] ...
Embodiment 2
[0023] The acquisition of Cryptosporidium-specific surface protein Cowp and the acquisition of quantum dot-labeled polystyrene fluorescent microspheres with a particle size of 4-6 μm coated with Cryptosporidium-specific surface protein Cowp:
[0024] For the gene sequence of Cowp, the primers we designed are:
[0025] Cowp-F:GCG GAATTC TAAATGTTTACATTTTCAGGGAAGCAG (SEQ ID. NO. 6)
[0026] Cowp-R:GCG CTCGAG TAACCTTGCAGTGTAAAATTTGGGG (SEQ ID. NO. 7)
[0027] Firstly, gene amplification was performed by PCR technology, and PCR products were recovered by OMEGA gel recovery kit. The amplified Cowp target fragment was ligated with the pMD18-T vector to generate the cloned plasmid pMD18-T-Cowp and identified. Afterwards, the target fragment of Cowp was ligated with the expression vector pTIG-Trx.
[0028] The method is to first extract the plasmid of the cloned plasmid pMD18-T-Cowp-positive strain, use EcoRI and XhoI double enzymes to cut the Cowp gene fragment, and recover and e...
Embodiment 3
[0031] The acquisition of Cryptosporidium-specific surface protein Cp41 and the acquisition of quantum dot-labeled polystyrene fluorescent microspheres with a particle size of 4-6 μm coated with Cryptosporidium-specific surface protein Cp41:
[0032] For the gene sequence of Cp41, the primers we designed are:
[0033] Cp41-F: TGGTGTTGCTTTTACTGCTATTCA (SEQ ID. NO. 8)
[0034] Cp41-R: AGTAGCAACAGTAGTAACAGTGG (SEQ ID. NO. 9)
[0035] Other operations were the same as in Example 2, and a large amount of Cp41 protein was obtained.
[0036] Coupling Cp41 protein with quantum dot-labeled polystyrene fluorescent microspheres with a particle size of 4-6 μm to obtain quantum dot-labeled polystyrene fluorescent microspheres with a particle size of 4-6 μm coated with Cryptosporidium-specific surface proteins Cp41.
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