Gholib Gholib, Wahyuni Sri, Akmal Muslim, Hasan Muhammad, Agil Muhammad, Purwantara Bambang
Physiology Laboratory, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Aceh, 23111, Indonesia.
Anatomy Laboratory, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Aceh, 23111, Indonesia.
F1000Res. 2019 Jul 30;8:1220. doi: 10.12688/f1000research.19804.3. eCollection 2019.
To obtain accurate measurements of cortisol (C) and testosterone (T) in Aceh cattle, commercial enzyme-linked immunosorbent assay (ELISA) kits need to be carefully validated. Moreover, repeated freeze-thaw cycles during the storage of the samples may affect the stability of the hormones in the serum. Here, the reliability of C and T concentration measurements in the serum of Aceh cattle, was tested using commercial C and T ELISA kits designed to measure human C and T concentrations. Further, the effect of repeated freeze-thaw cycles on the stability of C and T concentrations in the serum was evaluated. Commercial C (Cat. no. EIA-1887) and T (Cat. no. EIA-1559) ELISA kits from DRG Instruments GmbH were validated through an analytical validation test (i.e., parallelism, accuracy, and precision) and a biological validation test (for C: effect of transportation on the C secretion; for T: the concentrations of T between bulls and cows). To test the effects of freeze-thaw cycles, cattle serum was subjected to the following treatments: (i) remained frozen at -20 C (control group); (ii) exposed to freeze-thaw cycles for two, four, six, and eight times (test groups). Parallelism, accuracy, and precision tests showed that both C and T ELISA kits adequately measured C and T in the serum of Aceh cattle. Concentrations of C post-transportation were significantly higher than pre-transportation (p<0.01). Concentrations of T in bulls were significantly higher than in cows (p<0.01). After four to eight freeze-thaw cycles, C concentrations were significantly lower compared to the control group (all p < 0.05). In contrast, T concentrations remained stable (all p>0.05). Commercial C (EIA-1887) and T (EIA-1559) ELISA kits are reliable assays for measuring serum C and T, respectively, in Aceh cattle. Repeated freeze-thaw cycles significantly affected the stability of serum C, but did not for T.
为了准确测量亚齐牛的皮质醇（C）和睾酮（T），需要仔细验证商用酶联免疫吸附测定（ELISA）试剂盒。此外，样品储存期间反复的冻融循环可能会影响血清中激素的稳定性。在此，使用旨在测量人C和T浓度的商用C和T ELISA试剂盒，测试了亚齐牛血清中C和T浓度测量的可靠性。此外，还评估了反复冻融循环对血清中C和T浓度稳定性的影响。来自DRG Instruments GmbH的商用C（产品编号EIA - 1887）和T（产品编号EIA - 1559）ELISA试剂盒通过分析验证测试（即平行性、准确性和精密度）和生物学验证测试（对于C：运输对C分泌的影响；对于T：公牛和母牛之间T的浓度）进行了验证。为了测试冻融循环的影响，牛血清接受了以下处理：（i）在-20℃下保持冷冻（对照组）；（ii）暴露于冻融循环2次、4次、6次和8次（测试组）。平行性、准确性和精密度测试表明，C和T ELISA试剂盒均能充分测量亚齐牛血清中的C和T。运输后C的浓度显著高于运输前（p<0.01）。公牛中T的浓度显著高于母牛（p<0.01）。经过4至8次冻融循环后，C浓度与对照组相比显著降低（所有p<0.05）。相比之下，T浓度保持稳定（所有p>0.05）。商用C（EIA - 1887）和T（EIA - 1559）ELISA试剂盒分别是测量亚齐牛血清C和T的可靠测定方法。反复的冻融循环显著影响血清C的稳定性，但对T没有影响。
