Comparison of CLIA and ELFA Progesterone Assay Methods for Evaluating Pregnancy in Kilis Goats

Year 2025, Volume: 36 Issue: 3, 205 - 212, 30.11.2025

Nurdan Coşkun , Fikret Karaca Cafer Tayyar Ateş

https://doi.org/10.36483/vanvetj.1728594

Abstract

Early pregnancy diagnosis is needed for small ruminants. Effective methods for early pregnancy detection are of great importance. Identifying pregnancy enhances rebreeding and is crucial for controlled breeding programs. However, various methods exist for diagnosing pregnancy in small ruminants. The choice of methods is influenced by equipment availability, the number of days post-insemination and the required accuracy level. Measurement of progesterone concentrations is used in pregnancy diagnosis in small ruminants. The detection of progesterone (P4) assays can facilitate the diagnosis of pregnancy approximately three weeks post-insemination. This study aimed to evaluate and compare the accuracy of two progesterone assays (CLIA and ELFA) for the early diagnosis of pregnancy (at day 21 post-insemination) in Kilis goats, using transrectal ultrasonography at day 50 as the reference standard. Kilis goats (n: 150) were synchronized with a 10-day intravaginal sponge. Artificial insemination applications were intracervically performed with frozen-thawed semen straws. Blood samples were collected 21 days post-insemination to assess progesterone levels for pregnancy diagnosis. The pregnancy rate and average progesterone level determined by the CLIA method were found to be higher than those determined by the ELFA method. Additionally, in early pregnancy determination, the number of truly pregnant animals was significantly higher, with sensitivity, specificity, and accuracy. The CLIA method is a precise and dependable approach for measuring plasma progesterone levels. In the study, it was concluded that the CLIA progesterone assay method evaluated for early pregnancy detection in Kilis goats was superior.

Keywords

CLIA , ELFA , Goat , Pregnancy , Progesterone levels

Ethical Statement

The Hatay Mustafa Kemal University Scientific Ethical Committee approved the experiments (2022/08-01).

Kilis Keçilerinde Gebeliğin Değerlendirilmesinde CLIA ve ELFA Progesteron Analiz Yöntemlerinin Karşılaştırılması

Abstract

Küçük ruminantlarda erken gebelik tanısı gereklidir. Gebeliğin erken tespiti için etkili yöntemler büyük önem taşır. Gebeliğin belirlenmesi yeniden üremeyi artırır ve kontrollü üreme programları için çok önemlidir. Küçük ruminantlarda gebeliğin teşhisi için çeşitli yöntemler mevcut olsa da yöntem seçimi ekipman mevcudiyetine üreme sonrası gün sayısına ve gereken doğruluk düzeyine bağlıdır. Küçük ruminantlarda gebelik teşhisinde progesteron konsantrasyonlarının ölçümü kullanılır. Progesteron (P4) analizlerinin tespiti üreme sonrası yaklaşık üç hafta içinde gebelik teşhisini kolaylaştırabilir. Bu makale, CLIA ve ELFA yöntemleri ile gebelik teşhisinin hassasiyetini değerlendirmeyi önermektedir. Kilis keçileri (n:150) 10 günlük intravajinal süngerle senkronize edildi. Suni tohumlama uygulamaları dondurulmuş-çözülmüş semen payetleriyle intraservikal olarak gerçekleştirildi. Gebelik teşhisi için progesteron seviyelerini değerlendirmek amacıyla suni tohumlamadan 21 gün sonra kan örnekleri toplandı. CLIA yöntemi ile belirlenen gebelik oranı ve ortalama progesteron seviyesi ELFA yöntemi ile belirlenenlerden daha yüksek bulunmuştur. Ayrıca erken gebelik tespitinde gerçekten gebe hayvan sayısı duyarlılık, özgüllük ve doğruluk açısından önemli ölçüde daha yüksekti. CLIA yöntemi plazma progesteron seviyelerini ölçmek için hassas ve güvenilir bir yaklaşımdır. Çalışmada Kilis keçilerinde erken gebelik tespiti için değerlendirilen CLIA progesteron testi yönteminin üstün olduğu sonucuna varılmıştır.

Keywords

CLIA , ELFA , Gebelik , Keçi , Progesteron seviyeleri

References

• Al-janabi HSO, Kelkawi AHA, Abdul-Hsin IF, Kadhim MJ, Hussein MM (2018). Comparative evaluation study of ELISA system and MINI-VIDAS system for detection of Cytomegalovirus IgM antibodies. JPSR, 10 (10), 2549-2550.
• Al‐Merestani MR, Zarkawi M, Wardeh MF (2003). Improving the reproductive efficiency, pregnancy diagnosis and monitoring the resumption of luteal activity in indigenous Damascus goats. Reprod Domest Anim, 38 (1), 36-40.
• Alwan AF, Amin FAM, Ibrahim NS (2010). Blood progesterone and estrogen hormones level during pregnancy and after birth in Iraqi sheep and goat. Basra J Vet Res, 9 (2), 153-157.
• Amer HA (2010). Ultrasonographic assessment of early pregnancy diagnosis, fetometry and sex determination in goats. Anim Reprod Sci, 117 (3-4), 226–231.
• Arcot PJ, Pandey HC, Coshic P et al. (2022). Comparative evaluation of ADVIA Centaur® XP chemiluminescence system for screening of HBV, HCV, HIV and syphilis in Indian blood donors. Transfus Apher Sci, 61 (2), 103318.
• Bolet G (1986). Timing and extent of embryonic mortality in pigs sheep and goats: Genetic variability.
• Sreenan JM, Diskin MG (Eds.). Embryonic mortality in farm animals (pp. 34). Curr T Vet, Springer, Dordrecht.
• Boscos CM, Samartz FC, Lymber AG, Stefanakis A, Belibasaki S (2003). Assessment of progesterone concentration using enzymeimmunoassay for early pregnancy diagnosis in sheep and goats. Reprod Domest Anim, 38 (3), 170-174.
• Braun WF, Solorzano NM, Bierschwal CJ (1988). Characterization of the caprine estrous cycle using enzyme immunoassay for the determination of whole blood progesterone concentration. Theriogenology, 29 (5), 1155-1162.
• Brugger N, Otzdorff C, Walter B (2011). Quantitative determination of progesterone (P4) in canine blood serum using an enzyme-linked fluorescence assay. Reprod Domest Anim, 46 (5), 870-873.
• Capezzuto A, Chelini MOM, Felippe ECG, Olivera CA (2009). Correlation between serum and fecal concentration of reproductive steroids throughout gestation in goats. Anim Reprod Sci, 103 (1-2), 78-86.
• Chapwanya A, Clegg T, Stanley P (2008). Comparison of the Immulite and RIA assay methods for measuring peripheral blood progesterone levels in Greyhound bitches. Theriogenology, 70 (5), 795-799. Cinquanta L, Fontana DE, Bizzaro N (2017). Chemiluminescent immunoassay technology: what does it change in autoantibody detection? Autoimmun, 8 (1), 1-8. Dawson LJ (1999). Pregnancy diagnosis in goats. In: Proceedings of the 14th Annual Goat Field Day. Langston University, Langston.
• Dionysius DA (1991). Pregnancy diagnosis in dairy goats and cows using progesterone assay kits. Aust Vet J, 68 (1), 14-16.
• Doize F, Vaillancourt D, Carabin H, Belanger D (1997). Determination of gestational age in sheep and goats transrectal ultrasonographic measurement of placentome. Theriogenology, 48 (3), 449-460.
• Engeland IV, Ropstad E, Andresen Ø, Eik LO (1997). Pregnancy diagnosis in dairy goats using progesterone assay kits and oestrous observation. Anim Reprod Sci, 47 (3), 237-243.
• Fatet A, Pellicer-Rubio MT, Leboeuf B (2011). Reproductive cycle of goats. Anim Reprod Sci, 124 (3-4), 211-219.
• Fleming SA, Van Camp SD, Chapin HM (1990). Serum progesterone determination as an aid for pregnancy diagnosis in goats bred out of season. Can Vet J, 31 (2), 104-107.
• Fontbonne A, Maenhoudt C, Thoumire S et al. (2021). Evaluation of surface plasmon field-enhanced fluorescence spectroscopy for rapid measurement of progesterone concentration in bitches. Am J Vet Res, 82 (5), 417-424.
• Gaafar KM, Gabr MK, Teleb DF (2005). The hormonal profile during the estrous cycle and gestation in Damascus goats. Small Rumin Res, 57 (1), 85-93.
• Goel AK, Agrawal KP (1992). A review of pregnancy diagnosis techniques in sheep and goats. Small Rumin Res, 9 (3), 255-64.
• González F, Cabrera F, Batista M et al. (2004). A comparison of diagnosis of pregnancy in the goat via transrectal ultrasound scanning, progesterone, and pregnancy-associated glycoprotein assays. Theriogenology, 62 (6), 1108-1115.
• Gül S, Keskin M, Göçmez Z, Gündüz Z (2016). Effects of supplemental feeding on performance of Kilis goats kept on pasture condition. Ital J Anim Sci, 15 (1), 110-115.
• Haibel GK (1990). Use of ultrasonography in reproductive management of sheep and goat herds. Vet Clin North Am Food Anim Pract, 6 (3), 597-613.
• Hesselink JW, Taverne MAM (1994). Ultrasonography of the uterus of the goat. Vet Q, 16 (1), 41-45.
• Hollinshead FK, Hanlon DW (2019). Normal progesterone profiles during estrus in the bitch: A prospective analysis of 1420 estrous cycles. Theriogenology, 125, 37-42.
• Humblot P, De Montigny G, Jeanguyot N et al. (1990). Pregnancy-specific protein B and progesterone concentrations in French Alpine goats throughout gestation. Reproduction, 89 (1), 205-212.
• Ishwar AK (1995). Pregnancy diagnosis in sheep and goats: a review. Small Rumin Res, 17 (1), 37-44.
• Islam MM, Kizaki K, Takahashi T et al. (2014). Pregnancy diagnosis in Black Bengal goat by progesterone assay. Bangladesh J Anim Sci, 43 (3), 180-184.
• Jack AMM, Chang CC, Peh HI, Chan JPW (2012). Fecal progesterone for monitoring reproductive status in dairy goats. Turk J Vet Anim Sci, 36 (5), 566–572.
• Karadaev M (2015). Pregnancy diagnosis techniques in goats-a review. Bulg J Vet Med, 183 (3), 183-193.
• Khadiga MG, Mohamed KG, Doaa FT (2005). The hormonal profile during the estrous cycle and gestation in Damascus goats. Small Rumin Res, 57, 85-93.
• Khalifa EI (2015). Physiological response of zaraibi billy goats spermatozoa to soybean lecithin addition as an alternative to egg yolk varieties in semen extenders. Egypt J Anim Prod, 52 (3), 155-162.
• Kharche SD, Jindal SK, Priyadharsini R et al. (2013). Fertility following frozen semen artificial insemination in Jamunapari goats. Indian J Anim Sci, 83 (10), 1071-1073.
• Laiblin C (1991). Estrus detection in the bitch with a progesterone rapid test (EIA)- A sensible supplement to vaginal cytology. Tierärztl Prax, 19 (2), 197-199.
• Leboeuf B, Restall B, Salamon S (2000). Production and storage of goat semen for artificial insemination. Anim Reprod Sci, 62 (1-3), 113-141.
• Lemley CO, Camacho LE, Hallford DM, Vonnahme KA (2018). Uteroplacental secretion of progesterone and estradiol-17β in an ovine model of intrauterine growth restriction. Anim Reprod Sci, 193, 68-78.
• Martemucci G, D’Alessandro AG (2011). Induction/synchronization of oestrus and ovulation in dairy goats with different short term treatments and fixed time intrauterine or exocervical insemination system. Anim Reprod Sci, 126 (3-4), 187-194.
• Matsas D (2007). Pregnancy diagnosis in goats. Youngquist RS. Threlfall WR (Eds). Current Therapy in Large Animal Theriogenology (pp. 547-554), Saunders, Philadelphia.
• Medan М, Watanabe G, Absy G et al. (2004). Early pregnancy diagnosis by means of ultrasonography as a method of improving reproductive efficiency in goats. J Reprod Dev, 50 (4), 391-397. Moxon R, Copley D, England GC (2010). Technical and financial evaluation of assays for progesterone in canine practice in the UK. Vet Rec, 167 (14), 528-531.
• Nunes JF, Salgueiro CCM (2011). Strategies to improve the reproductive efficiency of goats in Brazil. Small Rumin Res, 98 (1-3), 176-184.
• Patton PE, Lim JY, Hickok LR et al. (2014). Precision of progesterone measurements with the use of automated immunoassay analyzers and the impact on clinical decisions for in vitro fertilization. Fertil Steril, 101 (6), 1629-1636.
• Pehlivan E, Polat H, Dellal G (2017). Annual change of reproductive hormones in female angora goats. TURJAF, 5 (4), 343-348.
• Proverbio D, Perego R, Spada E et al. (2013). Comparison of VIDAS and radioimmunoassay methods for measurement of cortisol concentration in bovine serum. Sci World J, 1-5.
• Russell NJ, Foster S, Clark P et al. (2007). Comparison of radioimmunoassay and chemiluminescent assay methods to estimate canine blood cortisol concentrations. Aust Vet J, 85 (12), 487-494. Tal S, Mazaki-Tovi M, Druker S et al. (2020). Evaluation of two chemiluminescent assays compared with radioimmunoassay for serum progesterone measurement in bitches. Theriogenology, 147, 116-123.
• Thibier M, Jeanguyot N, De Montigny G (1982). Accuracy of early pregnancy diagnosis in goats based on plasma and milk progesterone concentrations. Int Goat Sheep Res, 45 (2), 1-6.
• Thorburn GD, Schneider W (1972). The progesterone concentration in the plasma of the goat during the oestrous cycle and pregnancy. J Endocrinol, 52 (1), 23-36.
• Trabert B, Falk RT, Stanczyk FZ et al. (2015). Reproducibility of an assay to measure serum progesterone metabolites that may be related to breast cancer risk using liquid chromatography-tandem mass spectrometry. Horm Mol Biol Clin Investig, 23 (3), 79-84.
• Wani NA, Wani GM, Mufti M, Khan MZ (1998). Ultrasonic pregnancy diagnosis in gaddi goats. Small Rumin Res, 29 (2), 239-240.
• Williams CSF (1986). Pregnancy diagnosis in goat. Morrow DA (Ed). Current Therapy in Theriogenology (pp. 587-588), WB Saunders, Philadelphia.
• Yaganoglu AM, Topal M (2018). Morkaraman-İvesi ve Tuj Irkı koyunlarda gebelik testlerinin Roc analizi ile karşılaştırılması. Alinteri J Agric Sci, 33 (1), 29-36.
• Yalcin BC (1986). Sheep and goats in Turkey. FAO Anim Prod Health Pap, 60, 168.
• Yazici E, Ozenc E, Celik HA, Ucar M (2018). Ultrasonographic foetometry and maternal serum progesterone concentrations during pregnancy in Turkish Saanen goats. Anim Reprod Sci, 197, 93-105.
• Zamfirescu S, Anghel A, Nadolu D, Dobrin N (2011). Plasmatic profiles of pregnancy-associated glycoprotein and progesterone levels during early pregnancy in Carpathian Goat. Ann Rom Soc Cell Biol, 16 (2), 50-53.
• Zarkawi M (1997). Monitoring the reproductive performance in Awassi ewes using progesterone radioimmunoassay. Small Rumin Res, 26 (3), 291-294.
• Zarkawi M, Al-Masri MR (2002). Use of radioimmunoassay to measure progesterone levels during different reproductive stages in female Damascus goats. Trop Anim Health Prod, 34 (6), 535.