Detection of pregnancy associated protein(s) in the sera of pregnant buffaloes

Abstract

In the present study. attempt has been made to use enzyme linked immuno sorbent assay (ELISA) for the detection of
pregnancy-associated proteins in the scra of pregnant buffaloes. The blood samples from pregnant (n =25 . 30 days post insemination
till term) and non-pregnant buffaloes (n=l 0) were collected by jugular vein puncture and indirect plate test and dot ELISA was
performed with standard protocol using hyper immune sera raised against purified fractions of reproductive fluid from pregnant
genital tracts and placental extracts as a primary antibody solution . The optical density in indirect plate ELISA ranged from 0.515
to 0.878 (Mean 0.717), 0.327 to 0.710 (Mt:an 0.562) for pregnant samples and from 0.251 to 0.469 (Mean 0338), 0.209 to 0.350
(Mean 0.267) for non-pregnant sample using the anti- purified fraction of reproductive fluid and placental extract sera separately.
Indirect plate ELISA revealed higher OD values for pregnant scra sam ples indicating presrnce of some proteins associated with
pregnancy in the sera samples. In dot ELISA. out of 25 samples from pregnant buffaloes, positive color reaction was observed in 20
(80%) and I 7(68%) sera samples using anti-purified fraction of reproductive tluid and placental extract sera separately, however, no
color development was observed in 9(n= I 0. 90%) non pregnant samples . It appears that pregnancy associated proteins can be
detected in the sera of pregnant buffaloes further purification of these proteins mav enhance the accuracy.