Pretreatment-free lateral flow enzyme immunoassay for progesterone detection in whole cows’ milk

• Lateral flow enzyme immunoassay (LFEIA) for milk progesterone detection was developed.
• Horseradish peroxidase was used in combination with 3,3’,5,5’-tetramethylbenzidine.
• Pretreatment-free whole cows’ milk LFEIA can be performed within 15 minutes.
• Progesterone concentration determined by LFEIA and ELISA were in good correlation.

New rapid method of lateral flow enzyme immunoassay (LFEIA) for progesterone detection in whole cows’ milk was developed. The test system utilized horseradish peroxidase as a label along with the substrate solution containing 3,3’,5,5’-tetramethylbenzidine and dextran sulfate to obtain an insoluble blue colored product of the enzyme reaction on a surface of analytical membrane (test and control lines). Several aspects of LFEIA were optimized: time of the signal detection, membrane materials and assay conditions. Resulting competitive LFEIA can be performed within 15 minutes with the limit of progesterone detection of 0.8 ng/ml. Progesterone concentration in whole milk samples was determined by LFEIA and enzyme-linked immunosorbent assay (ELISA). The results obtained were in good correlation (R=0.97, n=46). Thus new sensitive LFEIA can be successfully used for on-site monitoring of oestrus status of cows’ reproductive system and for early none-pregnancy detection. The method is fast, easy to perform and needs no preliminary sample preparation.

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