Analytical validation of an enzyme‐linked immunosorbent assay for the quantification of S100A12 in the serum and feces of cats

2019 ◽  
Vol 48 (4) ◽  
pp. 754-761
Author(s):  
Cory S. Bridges ◽  
Niels Grützner ◽  
Jan S. Suchodolski ◽  
Jörg M. Steiner ◽  
Romy M. Heilmann
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Validation

scholarly journals The validation of a commercial enzyme-linked immunosorbent assay and the effect of freeze-thaw cycles of serum on the stability of cortisol and testosterone concentrations in Aceh cattle

F1000Research ◽  
2020 ◽  
Vol 8 ◽  
pp. 1220
Author(s):  
Gholib Gholib ◽  
Sri Wahyuni ◽  
Muslim Akmal ◽  
Muhammad Hasan ◽  
Muhammad Agil ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Analytical Validation ◽  
Commercial Enzyme ◽  
Freeze Thaw ◽  
Validation Test ◽  
Accuracy And Precision ◽  
Cattle Serum ◽  
The Stability

Background: 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. Methods: 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 -20OC (control group); (ii) exposed to freeze-thaw cycles for two, four, six, and eight times (test groups).   Results: 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). Conclusions: 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.

scholarly journals The validation of a commercial enzyme-linked immunosorbent assay and the effect of freeze-thaw cycles of serum on the stability of cortisol and testosterone concentrations in Aceh cattle

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 1220 ◽  
Author(s):  
Gholib Gholib ◽  
Sri Wahyuni ◽  
Muslim Akmal ◽  
Muhammad Hasan ◽  
Muhammad Agil ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Analytical Validation ◽  
Commercial Enzyme ◽  
Freeze Thaw ◽  
Validation Test ◽  
Accuracy And Precision ◽  
Cattle Serum ◽  
The Stability

Background: 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, we test the reliability of C and T concentration measurements in the serum of Aceh cattle, obtained using commercial C and T ELISA kits designed to measure human C and T concentrations. Further, we evaluate the effect of repeated freeze-thaw cycles on the stability of C and T concentrations in the serum. Methods: 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 excretion; 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 -20OC (control group); (ii) exposed to freeze-thaw cycles for two, four, six, and eight times (test groups).   Results: 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<0.05). Concentrations of T in bulls were significantly higher than in cows (p<0.05). 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). Conclusions: 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.

scholarly journals MEASUREMENT OF SERUM TESTOSTERONE IN KACANG GOAT BYUSING ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) TECHNIQUE: THE IMPORTANCE OF KIT VALIDATION

2016 ◽  
Vol 10 (1) ◽  
Author(s):  
Gholib G ◽  
Sri Wahyuni ◽  
Okta Hilda Kadar ◽  
Mulyadi Adam ◽  
Triva Murtina Lubis ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Jugular Vein ◽  
Serum Testosterone ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Validation ◽  
Validation Data ◽  
Testosterone Concentration ◽  
Standard Curve ◽  
Biological Validation ◽  
Elisa Technique

This study was conducted to validate a commercial testosterone enzyme-linked immunosorbent assay (ELISA) kits (DRG EIA-1559) inanalytic and biological manner for measuring serum testosterone concentrations in kacang goats. This study used 18 healthy kacang goats, sixbucks (>2 years), six kids (<6 months), and six does (>2 years). Blood samples were collected from jugular vein and prepared as serum. Twovalidation tests were performed, an analytical validation comprises a parallelism, accuracy, precision and sensitivity and a biological validationby comparing testosterone concentration from bucks, kids, and does. Testosterone concentrations were measured using ELISA technique. Data ofanalytical validation were analyzed descriptively and test of equality of slope was performed to see the parallelism between samples and standardcurves. Analysis of variance (ANOVA) was used for biological validation data. Results of parallelism showed that sample curve was parallel tothe standard curve. Accuracy, precision (% CV of intra-and inter-assay) and sensitivity of the assay were 99.65±4.27%, <10%, <15% and 0.083ng/ml, respectively. Results of biological validation showed that the assay used were accurately measured testosterone which testosteroneconcentrations in bucks were significantly higher compared to kids and does (P<0.05). In conclusion, a commercial testosterone ELISA kits(DRG EIA-1559) is a reliable assay for measuring serum testosterone concentration in kacang goats. Key words: analytical and biological validations, ELISA, testosterone, kacang goat

scholarly journals The validation of a commercial enzyme-linked immunosorbent assay and the effect of freeze-thaw cycles of serum on the stability of cortisol and testosterone concentrations in Aceh cattle

F1000Research ◽  
2020 ◽  
Vol 8 ◽  
pp. 1220
Author(s):  
Gholib Gholib ◽  
Sri Wahyuni ◽  
Muslim Akmal ◽  
Muhammad Hasan ◽  
Muhammad Agil ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Analytical Validation ◽  
Commercial Enzyme ◽  
Freeze Thaw ◽  
Validation Test ◽  
Accuracy And Precision ◽  
Cattle Serum ◽  
The Stability

Background: 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. Methods: 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 excretion; 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 -20OC (control group); (ii) exposed to freeze-thaw cycles for two, four, six, and eight times (test groups).   Results: 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<0.05). Concentrations of T in bulls were significantly higher than in cows (p<0.05). 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). Conclusions: 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.

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