Validation Of Commercial Eia Kit For Analysis Of Hormone Estradiol And Progesterone in Female Kacang Goat (Capra hircus)

Abstract

Enzyme immunosorbent assay (EIA) is a technique that connects the specificity of antibodies with the enzymatic test sensitivity or antigen attached to the enzyme by regular spectrophotometer. Analysis using EIA technique has been proved suitable to replace the radio immunoassay (RIA) technique which has many weaknesses. There are several commercial EIA hormone kit products that can be used nowadays to test the concentration of the hormone, but not any of the products which can be used as a reference. The aims of this study was to determine the feasibility of commercial EIA hormone kits for human estradiol and progesterone whether can be used or not for monitoring reproductive status of kacang goat. This study used 3 female kacang goats 2–3 years old, healthy, have regular estrous cycles, fertile and unpregnant. Blood samples were taken from the jugular vein using a 21 G venoject every two days and sample collection was intensified every day prior to heat. Blood plasma stored at -20ºC until the analyse. Ultrasound is used to check the female reproductive organs (ovaries) during blood sampling. Hormone assay validation was conducted through laboratory validation and biologycal validation. Laboratory validation was carried out by measuring accuracy, sensitivity and precision. Comparison between hormone profile and morphologycal change and USG picture of the ovary was used as biologycal validation. Parallelism test showed that sample curve was parallel with standard curve of E2 and P4 of DRG commercial kit, in contrast GBC commercial kit was not parallel. The lowest hormone concentration of estradiol (E2) and progesterone (P4) at 90% binding was 25 pg/mL and 0.14 ng/mL in DRG kit. Coefficient variation of intra- and interassay for both DRG EIA commercial kits were less than 10%. Goat 5 and 9 did not show estrus cycle with hormone profiles appear flat. Goat 7 showed irregular estrous cycles, with only finding of progesterone profile of one cycle during 62 days of observation, although it did not coincide with profiles of estradiol on the cycle. Progesterone concentrations during the luteal phase ranged from 3.6-42.8 ng/mL. Concentration appears to increase significantly at day 4 after ovulation was observed and reached a peak on day 6 to day 14 with a concentration of 19.2-42.8 ng / mL before decreased dramatically by the end of the cycle. It can be concluded that the P4 DRG EIA kit is suitable and can be used for monitoring the reproductive status by measuring plasma samples female kacang goat, but not compatible for E2 EIA DRG. While E2 and P4 EIA kits GBC can not be used to analyze hormones E2 and P4 from blood samples of kacang goat.