Determination of progesterone concentration during the estrous cycle in dairy cows using a chemiluminescence assay

Kozicki, Luiz Ernandes; Weber, Saulo Henrique; Bacher, Louise Helene; Ramos, Isabella Sellmer; Gaievski, Francisco Romano; Schaus, Marcos; Talini, Rafaela; Weiss, Romildo Romualdo; Bergstein-Galan, Tácia Gomes; Segui, Marcio Saporski; Aita, Carlos Alberto Mayora

doi:10.7213/1981-4178.2018.16006

Cited by 1 publication

(5 citation statements)

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“…Therefore, it was observed that the P4 profile of the animals was consistent with the findings of follicular dynamics corroborated by Ginther et al (2007). With the data adjusted using the FCC, we observed that the serum P4 concentration patterns in mares obtained using CLIA was similar to those reported by Kozicki et al (2018) (value < 5.48 ng/mL of serum P4 defined the follicular phase and ≥ 5.48 ng/mL defined the luteal phase of the oestrous cycle). Therefore, it is possible to observe an increase in serum P4 in mares from d3 of the cycle extending to d17 (Tab.…”

Section: Discussionsupporting

confidence: 84%

“…The CLIA was proved to be a useful methodology capable of determining the P4 profile of dairy cows during the oestrous cycle. Kozicki et al (2018) demonstrated that a serum P4 value < 5.48 ng/ mL is indicative of the follicular phase and ≥ 5.48 ng/mL of the luteal phase in cows, which was further confirmed by following the oestrous cycle upon daily examination of ovarian ultrasound (US).…”

Section: Introductionmentioning

confidence: 85%

“…More recently, CLIA has become widely used owing to its acceptance and applicability in human medicine. It is gradually gaining attention among researchers from other research areas, especially veterinary medicine, regarding canines (Tahir et al 2013) and cows (Kozicki et al 2018).…”

Section: Discussionmentioning

confidence: 99%

“…Limited reports related to serum P4 determination in large animals using the CLIA methodology have been found. Kozicki et al (2018) determined P4 levels during the oestrous cycle in dairy cows. Few studies have been conducted to quantify P4 in mares using the CLIA methodology.…”

Section: Discussionmentioning

confidence: 99%

“…A more recent study was conducted in dairy cows, indicating that CLIA is a methodology capable of determining the profile of serum P4 in cows. In this research, the values < 5.48 ng/mL of serum P4 were established to define the follicular phase, and ≥ 5.48 ng/mL for the luteal phase of the oestrous cycle (Kozicki et al 2018).…”

Section: Discussionmentioning

confidence: 99%

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Serum progesterone profile in the oestrous cycle of mares determined using immuno chemiluminescence (CLIA)

Prus¹,

Kozicki²,

Weber³

et al. 2022

PHK

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Six cross-bred mares (Mangalarga, Quarter Horse, Apaloosa, Criollo) were used to (1) quantify the serum progesterone (P4) levels during the oestrous cycle of mares using chemiluminescence (CLIA), (2) compare the P4 profile quantified using CLIA and the follicular dynamics data obtained using ovarian ultrasound (US), and (3) validate the CLIA methodology with an enzyme-linked immunosorbent assay (ELISA). The horses were between 10 and 18 years old, body weight ranging from 350 to 490 kg, and with a body condition score of 3.5. We conducted the study starting from the day of ovulation until the next ovulation was detected using US examinations. Blood samples were collected at intervals of two days and stored at -20 °C for the determination of P4 using ELISA and CLIA. Concomitantly, the ovaries were scanned daily using US to monitor the follicular dynamics. The length of the oestrous cycle (interval ovulation to ovulation) varied between 22 and 27 days. The results of the P4 concentration were analysed using Pearson's χ² and Kolmogorov-Smirnov tests. The P4 concentration in the follicular phase ranged from 0.38 to 1.91 ng/mL (CLIA) and 0.81 to 3.26 ng/mL (ELISA), and the concentrations in the luteal phase were 8.69 to 29.05 ng/mL and 11.84 to 25.16 ng/mL, respectively. The P4 concentration ≥ 5.48 ng/mL was indicative of the luteal phase (days 3 to 15 of the cycle) and < 5.47 marked the follicular phase of the oestrous cycle (day 16 until the next ovulation). The dimensions (diameter) of the largest follicle and corpus luteum varied between 35.0 and 47.5 mm and 14.2 and 43.7 mm, respectively. A follicular wave was observed during the oestrous cycle of the animals. The follicular and luteal phases of the oestrous cycle were established based on P4 concentrations after the analysis using the χ² and Kolmogorov-Smirnov adherence tests. The Pearson's χ² test results indicated a p-value < 0.05, presenting different frequencies and numerical values between ELISA and CLIA; however, the curves of both the analyses depicted the same trend. Therefore, after adjustment, the Kolmogorov-Smirnov adherence test (KS at 95 % probability) was performed where the maximum difference between the observed and estimated frequencies was compared and divided by the number of observations. The values proved to be adherent, with the same tendency in the resulting curve. The CLIA proved to be an efficient method for determining the serum P4 in mares, as it determined the P4 profile, showing adherence to the ovarian findings verified using US examinations. Therefore, CLIA can be used as an alternative method because it has been proved to be an efficient, low-cost and rapid method.

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Section: Discussionsupporting

confidence: 84%

Section: Introductionmentioning

confidence: 85%