Development of equine chorionic gonadotrophin (eCG) based sandwich ELISA for pregnancy diagnosis and fetus viability in mares

ASHOK KUMAR GUPTA; YASH PAL; SANJAY KUMAR

doi:10.56093/ijans.v86i3.56647

Authors

ASHOK KUMAR GUPTA National Research Centre on Equines, Hisar, Haryana 125 001 India
YASH PAL National Research Centre on Equines, Hisar, Haryana 125 001 India
SANJAY KUMAR National Research Centre on Equines, Hisar, Haryana 125 001 India

https://doi.org/10.56093/ijans.v86i3.56647

Keywords:

eCG quantification, Fetus viability, Pregnancy diagnosis, Sandwich ELISA, Sensitivity, Specificity

Abstract

An equine chorionic gonadotrophin (eCG) based sandwich ELISA (sELISA) was developed for confirmation of early conception in pregnant mares around 30 days of gestation and fetus viability thereafter. This serum based quantitative assay is quite sensitive, specific, precise and efficient. Serum eCG content higher than 10 IU/ml during early gestation was used for confirmation of conception while eCG content more than 40 IU/ml was taken as cut off for fetus viability between 40 – 60 days of gestation. Assay was performed with more than 4,000 field serum samples and compared with equivalent commercial pregnancy diagnostic kit with its better sensitivity. Assay showed little cross reactivity with serum of non pregnant mares, jennies, and other animal species. Confirmation of fetus viability sELISA helped in re-servicing the mares which are observed to empty after conception failure or fetal loss.

Downloads

Download data is not yet available.

References

Alexander B M, Johnson M S, Guardia R O, Van de Graaf W L, Senger P L, and Sasser R G. 1995. Embryonic loss from 30 to 60 days post breeding and the effect of palpation per rectum on pregnancy. Theriogenology 43: 551–56. DOI: https://doi.org/10.1016/0093-691X(94)00060-8

Allen W R, Hamilton D W and Moor R M. 1973. The origin of equine endometrial cups. Invasion of the endometrium by trophoblast. Anatomical Record 177: 485–502. DOI: https://doi.org/10.1002/ar.1091770403

Allen W R and Moor R M. 1972. The origin of the equine endometrial cups. I. Production of PMSG by fetal trophoblast cells. Journal of Reproduction and Fertility 29: 313–16. DOI: https://doi.org/10.1530/jrf.0.0290313

Allen W R. 1969. The immunological measurement of pregnant mare serum gonadotrophin. Journal of Endocrinology 43: 593– 98. DOI: https://doi.org/10.1677/joe.0.0430593

Allen W R. 1975. The influence of fetal genotype upon endometrial cup development and PMSG and progestagen production in equids. Journal of Reproduction and Fertility Suppl 23: 405–13.

Allen W R. 1978. Control of Ovulation and Oestrus in Mare. Control of Ovulation. p. 453–70. (Eds) Crighton D P, Haynes H B, Foxcroft G R, Lamming G E. Butterworths, London DOI: https://doi.org/10.1016/B978-0-408-70924-8.50032-3

Clegg M T, Boda J M and Cole H H. 1954. The endometrial cups and allantochorionic pouches in the mare with emphasis on the source of equine gonadotrophin. Endocrinology 54: 448– 63. DOI: https://doi.org/10.1210/endo-54-4-448

Ginther O J, Garcia M C, Bergfelt D R, Leith G S and Scraba S T. 1985. Embryonic loss in mares: pregnancy rate, length of interovulatory intervals and progesterone concentrations associated with loss during days 11 to 15. Theriogenology 24: 409–17. DOI: https://doi.org/10.1016/0093-691X(85)90047-0

Harlow Ed and Lane D. 1988. Antibodies a Laboratory Manual. Cold Spring Harbor Laboratory, New York, USA.

Lewis L D. 1990. Broodmare feeding and care. Equine Clinical Nutrition: Feeding and Care. Lea and Febiger, Philadelphia p. 286–306.

Manning A W, Kumar R, Bristol F, Flood P F and Murphy B D. 1987. Genetic and temporal variation in serum concentrations and biological activity of horse chorionic gonadotrophin. Journal of Reproduction and Fertility Suppl 35: 389–97.

Martinuk S D, Bristol F and Murphy B D. 1990. Effects of the dam on equine chorionic gonadotrophin concentrations during pregnancy. Domestic Animal Endocrinology 7: 551–57. DOI: https://doi.org/10.1016/0739-7240(90)90012-O

Papa F O, Lopes M D, Alvarenga M A, De Meira C and Luvizotto M C R. 1998. Early embryonic death in mares: clinical and hormonal aspects. Brazilian Journal of Veterinary Research and Animal Science 35: 170–73. DOI: https://doi.org/10.1590/S1413-95961998000400005

Papkoff H. 1974. Chemical and biological properties of the subunits of pregnant mare serum gonadotrophin. Biochemical and Biophysical Research Communications 58: 397–404. DOI: https://doi.org/10.1016/0006-291X(74)90378-7

Rathwell A C, Asbury A C, Hansen P J and Archbald L F. 1987. Reproductive function of mares given daily injections of Prostaglandin F2 beginning at day 42 of pregnancy. Theriogenology 27: 621–30. DOI: https://doi.org/10.1016/0093-691X(87)90056-2

Relan M. 2001. ‘Studies on equine chorionic gonadotrophin (eCG) and development of immunoassay for pregnancy diagnosis in mares.’ Ph.D. Thesis, Department of Veterinary Physiology, CCS HAU, Hisar, India.

Richter W. 1963. Interruption of pregnancy in mares. Zuchtungskunde 7: 81–85.

Roberts S J. 1986. Veterinary Obstetrics and Genital Diseases. 2nd edn. New York: Woodstock, Ithaca.

Tyler J W and Cullor J S. 1989. Titres, tests and truism : rational interpretation of diagnostic serological testing. Journal of the American Veterinary Medical Association 11 : 1550–58.

Van Niekerk C H and Morgenthal J C. 1982. Fetal loss and the effect of stress on plasma progestagen levels in pregnant Thoroughbred mares. Journal of Reproduction and Fertility Suppl 32: 453–57.

Van Niekerk C H. 1965. The early diagnosis of pregnancy, the development of the foetal membranes and nidation in the mare. Journal of the South African Veterinary Medical Association 36: 483–88.

Virmani M, Gupta A K and Garg S K. 2006. Determination of gestation period of mare with maximum eCG concentration by bioassay. Indian Journal of Animal Research 40: 25–29.