An enzyme-linked immunosorbent assay (ELISA) to measure growth hormone level in serum and milk of buffaloes (Bubalus bubalis).
An indirect Sandwich ELISA to measure growth hormone level in serum and milk of buffaloes was developed. The assay was based on purified anti rbST IgG raised in rabbits and chicken and rabbit anti chicken IgG horseradish peroxidase. The assay was validated in terms of sensitivity, specificity, precision and recovery. Parallelism was demonstrated between the standard curve and serially diluted serum, milk and pituitary derived growth hormone. Sensitivity of the assay was 0.1 ng/ml. Recovery of exogenous bovine somatotropin from serum and milk ranged from 90 to 102% and 96 to 108% respectively. The intra and inter assay variations to measure growth hormone in serum and milk were 3.36 to 8.81% and 6.01 to 12.31% respectively. Statistical analysis for parallelism and cross-reactivity of rbST with serum of other species confirmed the reproducibility of the assay.
Mishra
A
A
Division of Physiology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243 122, India. ashishvet1@rediffmail.com
Goswami
T K
TK
Shukla
D C
DC
D000818:Animals; D002020:Buffaloes / Q000378:metabolism*; D002138:Calibration; D002417:Cattle; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods*; D005799:Genes, Dominant; D013006:Growth Hormone / Q000097:blood* / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry*; D015233:Models, Statistical; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity