Immunochromatographic test strip for rapid detection of ancylostomiasis infection and preparation method thereof
An immunochromatography and hookworm disease technology, which is applied in the field of bioengineering, can solve the problems of high requirements for instruments, equipment and operators, and is not suitable for on-site use, and achieves the effects of easy industrial production, low cost, and simple detection methods.
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Embodiment 1
[0038] Embodiment 1, the preparation of the immunochromatographic test strip that detects hookworm infection
[0039] 1. Preparation of purified antigen against hookworm
[0040] About 2000 adult hookworms of American hookworm were collected, washed repeatedly with phosphate buffer, and the worms were placed in a pre-cooled mortar and continuously poured into liquid nitrogen for thorough grinding. Then add 20 times the volume of worm body volume containing 1.5% n-octyl glucoside, 1mM phenylmethylsulfonyl fluoride, 0.2mM tosyllysyl chloromethanone, 1mM ethylenediaminetetraacetic acid, 1mM ethylene glycol bismuth (2-Aminoethyl ether) tetraacetic acid soaked and rinsed the collected parasite material, repeated freezing and thawing 5 times in liquid nitrogen and 37°C, and ice-bathed overnight. Centrifuge at 26000g at 4°C for 30 minutes, and dialyze the supernatant against phosphate buffer for 48 hours. Ammonium sulfate was added to the solution to 70% saturation and a precipitat...
Embodiment 2
[0061] Embodiment 2, the preparation of the immunochromatographic test strip that detects hookworm infection
[0062] Colloidal gold-labeled detection probes were prepared from Staphylococcus aureus protein A (SPA) purchased from Shanghai Yeli Biotechnology Co., Ltd., and the quality control line was coated with chicken anti-Staphylococcus aureus protein A IgY purchased from Jiening Biotechnology Co., Ltd. , other steps are with embodiment 1.
Embodiment 3
[0063] Embodiment 3, laboratory assessment of the immunochromatographic test strip for detecting hookworm infection
[0064] 1. Detection method
[0065] Add the serum to be tested to the blood filtration membrane sample pad of the immunochromatography test strip prepared in Example 1, add 1 drop (about 50 μL) of sample diluent (PBS) after 1 minute, start to observe the results after 2 minutes, and stop after 20 minutes . Judgment of results: If red bands appear on both test line 5 and quality control line 6, it is judged as hookworm disease; if only red bands appear on quality control line 6, it is judged as negative.
[0066] 2. Experimental results
[0067] Table 1 shows the results of laboratory assessment using the immunochromatographic immunochromatographic test strip for detecting hookworm infection prepared in Example 2 of the present invention. It can be seen from Table 1 that the sensitivity of the immunochromatographic test paper of the present invention can reac...
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