I am looking for the supplier of an ELISA kit for the diagnosis of human fasciolosis, using coproantiges.
I found this article that maybe of help.
A Coproantigen Diagnostic Test for Strongyloides Infection
by Alex M. Sykes, James S. McCarthy
In their methodology they describe an ELISA used
Coproantigen ELISA
The assay components were titrated by doubling dilution to determine optimal signal:noise ratios. 96 well flat bottom microtiter plates (Nunc, Thermo Scientific) were coated with 50 µl (5 µg/ml) α-E/S Ig diluted in coating buffer (15 mM Na2CO3, 35 mM NaHCO3, pH 9.6) and incubated overnight at 4°C or at room temperature for 90 min. After incubation, wells were washed 4–5 times in PBS-T (PBS containing 0.05% Tween20). Wells were then blocked with 150 µl of PBS containing 2% casein (Merck, San Diego, CA) for 60 minutes at room temperature. Uninfected rat fecal supernatant (nRFS) and infected fecal supernatants (iRFS) were diluted 1:4 in PBS unless otherwise specified, and 50 µl added to each well and incubated for 90 min at room temperature. Known positive and negative formalin-fixed rat fecal supernatants, E/S products diluted in PBS and uninfected formalin-fixed fecal supernatant were assayed in duplicate. Wells containing only PBS or nRFS were included on each plate for standardization. Plates were then washed 4 times in PBS-T, after which 50 µl of 1:500 α-E/S-B was added, and incubated at room temperature for 60 min. The plates were again washed 4 times in PBS-T and 50 µl of NeutrAvidin-HRP (Thermo Scientific) diluted 1:10,000 was added, and the plates incubated for a further 60 min at room temperature. Plates were then washed 4 times in PBS-T prior to the addition of 100 µl per well of the developing substrate, ABTS (2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonicacid)). After incubation at room temperature for 20–30 min plates were read on a Versamax microplate reader (Molecular Devices) at 405 nm, and analysed with Prism 4 (Graphpad software, La Jolla, CA).
Hello there,
I am aware of Fasciola hepatica copro antigen ELISA kit made by Bio K 201 in sheep. Full info available here: http://search.cosmobio.co.jp/cosmo_search_p/search_gate2/docs/BOX_/BIOK2012.20120926.pdf.
I am not sure if the kit has been validated for use in Humans but you can contact them for information. I hope that helps.
Nusi
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