title: Pretreatment-free lateral flow enzyme immunoassay for progesterone detection in whole cows' milk.
abstract: 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.
authors.affiliation: Department of Chemical Enzymology, Chemistry Faculty, M.V. Lomonosov Moscow State University, 1-11 Leninskie Gory, Moscow, 119991 Russia. Electronic address: jvs@enz.chem.msu.ru.
authors.forename: J V
authors.initials: JV
authors.lastname: Samsonova
authors.affiliation: Department of Chemical Enzymology, Chemistry Faculty, M.V. Lomonosov Moscow State University, 1-11 Leninskie Gory, Moscow, 119991 Russia. Electronic address: capreace@inbox.ru.
authors.forename: V A
authors.initials: VA
authors.lastname: Safronova
authors.affiliation: National University of Science and Technology "MISiS", 4 Leninsky Ave., Moscow, 119049 Russia. Electronic address: aposipov@mail.ru.
authors.forename: A P
authors.initials: AP
authors.lastname: Osipov
mesh_terms: D000818:Animals; D001560:Benzidines / Q000737:chemistry; D016264:Dextran Sulfate / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry; D007124:Immunoenzyme Techniques / Q000295:instrumentation / Q000379:methods*; D057230:Limit of Detection; D008892:Milk / Q000737:chemistry*; D011374:Progesterone / Q000032:analysis*
chemical_list: D001560:Benzidines; C021758:3,3',5,5'-tetramethylbenzidine; D011374:Progesterone; D016264:Dextran Sulfate; D006735:Horseradish Peroxidase
keywords: Horseradish peroxidase; Lateral flow enzyme immunoassay; Whole cows’ milk; Рrogesterone