Rapid flow-through enzyme immunoassay of progesterone in whole cows' milk

Anal Biochem. 2018 Mar 15:545:43-48. doi: 10.1016/j.ab.2018.01.011. Epub 2018 Jan 31.

Abstract

A rapid flow-through immunoassay using an enzyme (horseradish peroxidase) as a label for quantitative and semi-quantitative determination of progesterone in whole cows' milk was developed. The flow-through test device consisted of a porous nitrocellulose membrane coated with antibodies and an absorbent membrane. The substrate solution containing 3,3',5,5' -tetramethylbenzidine was used for colour visualization. The detection limit of 0.4 ng/mL P4 was obtained by this method; analysis time did not exceed 15 min. To eliminate matrix interference a simple sample preparation procedure was used. Results of analysis of whole cows' milk samples with flow-through method were in good correlation with ELISA results (R = 0.96, n = 34). The developed rapid flow-through test system showed high efficiency for the determination of progesterone level in whole cow's milk and can be used on-site for quick identification of milk samples with low and high progesterone concentration.

Keywords: Flow-through enzyme immunoassay; Horseradish peroxidase; Progesterone; Whole cows' milk.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzidines / chemistry
  • Benzidines / metabolism
  • Cattle
  • Female
  • Horseradish Peroxidase / metabolism*
  • Immunoenzyme Techniques
  • Milk / chemistry*
  • Milk / metabolism
  • Progesterone / analysis*
  • Progesterone / metabolism

Substances

  • Benzidines
  • 3,3',5,5'-tetramethylbenzidine
  • Progesterone
  • Horseradish Peroxidase