ORBi: Delahaut Ph - Development of a new ELISA for measurement of pregnancy-associated glycoprotein.

Reference : Development of a new ELISA for measurement of pregnancy-associated glycoprotein.


	Document type :	Scientific congresses and symposiums : Paper published in a book
	Discipline(s) :	Life sciences : Veterinary medicine & animal health
	To cite this reference:	http://hdl.handle.net/2268/103766

Title :	Development of a new ELISA for measurement of pregnancy-associated glycoprotein.
Language :	English
Author, co-author :	Delahaut, Ph [ > > ]
	Melo de Sousa, Noelita [Université de Liège - ULg > Département de sciences fonctionnelles > Physiologie de la reproduction >]
	Huet, A. C. [ > > ]
	Colemonts, Y. [ > > ]
	Bertrand, Henri [Université de Liège - ULg > > > 3e an.méd. vété.]
	Barbato, O. [ > > ]
	Beckers, Jean-François [Université de Liège - ULg > Département de sciences fonctionnelles > Physiologie de la reproduction >]
Publication date :	7-Sep-2011
Main document title :	Proceedings of the 6th European Congress of Bovine Health and Management (ECBHM)
Pages :	95
Peer reviewed :	No
On invitation :	No
Audience :	International
ISBN :	9789079892020
Country :	Belgium
Event name :	6th European Congress of Bovine Health and Management (ECBHM)
Event date :	7-9 septembre 2011
Keywords :	[en] Pregnancy-associated glycoprotein ; ELISA ; development ; cattle ; pregnancy
Abstract :	[en] In the present study we report the development of a new ELISA for PAG measurement (Ref. code E.G.7, CER, Maloie) in ruminant species. Purified boPAG67kDa protein was purified in our laboratory and used as standard. Antiserum raised against caprine PAG 55kDa+62kDa (AS707) was used as capture antibody (1:40,000). Antiserum raised against PAG purified from buffalo placentas (AS859) was used as detection antibody (1:32,000 dilution). Avidin-HRP and TMB completed the ELISA system. Reproducibility was determined by calculating the intra- and inter-assay CV. Accuracy was determined by adding increasing concentrations of purified boPAG67kDa to bovine sera containing low PAG concentrations. Specificity was tested regarding 25 different compounds: carbohydrates, aspartic proteinases, hormones and major plasma proteins. Parallelism was assessed by serially diluting pregnant cow serum containing high PAG concentrations. Sera was collected from pregnant cows (n=4) during the whole duration of gestation. Samples were tested by both RIA and ELISA. Samples giving high PAG concentrations were serially diluted in order to fit with optical density (OD) of the ELISA standard curve. Intra-and inter-assay CV ranged from 5.5 to 9.1%. Accuracy ranged from 92.9 to 105.5%. No cross reaction was observed with almost all tested compounds excepting lactic acid at supra-physiological levels (1 mg/ml). Dilutions of bovine maternal sera were parallel to standard curve. Profiles obtained by ELISA in pregnant cows were very similar to those obtained by RIA. In conclusion, a new PAG-ELISA is available for measurement of PAG concentrations in cattle. This new test was validated in vitro and can be used for pregnancy diagnosis and follow-up of gestation in cattle.
Target :	Researchers ; Professionals ; Students
Permalink :	http://hdl.handle.net/2268/103766

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	File	Commentary	Version	Size	Access
Restricted access	DelahautPh_6ECBHM_2011.pdf		Publisher postprint	199.59 kB	Request copy
Restricted access	114e_ECBHM_ELISA_240811.pdf		Publisher postprint	323.56 kB	Request copy

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