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Validation and determination of a reference interval for canine HbA1c using an immunoturbidimetric assay
Anne F Goemans1, Susanna J Spence1, Ian K Ramsey1
1Small Animal Hospital, University of Glasgow, Glasgow, UK.
Insights
This study validates a new assay for measuring Hemoglobin A1c (HbA1c) in dogs, establishing a reference interval for this important diabetes mellitus marker. The validated assay offers reliable canine HbA1c measurement for improved glycemic control monitoring.
Area of Science:
- Veterinary Medicine
- Clinical Pathology
- Endocrinology
Background:
- Hemoglobin A1c (HbA1c) is a key indicator of long-term glycemic control in humans.
- While HbA1c exists in dogs, validated commercial assays for its measurement are lacking.
- Accurate HbA1c measurement is crucial for managing canine diabetes mellitus.
Purpose of the Study:
- To validate a commercially available automated immunoturbidimetric assay for canine HbA1c.
- To establish a reference interval (RI) for canine HbA1c in a hospital population.
- To assess the analytical performance of the assay for canine use.
Main Methods:
- Assay validation included specificity, repeatability, long-term imprecision, stability, and linearity testing.
- The effect of common anticoagulants (EDTA, heparin) on HbA1c measurement was evaluated.
- A reference interval was determined using blood samples from 60 healthy hospitalized dogs.
Main Results:
- The assay demonstrated proportional response to glucose concentration in vitro.
- Repeatability (CV 4.08%) and stability (5 days at 4°C) were confirmed.
- The reference interval for canine HbA1c was determined to be 9-18.5 mmol/mol, with no significant effect of age or breed.
Conclusions:
- The validated automated immunoturbidimetric assay provides reliable and analytically sound measurement of canine HbA1c.
- This assay facilitates improved monitoring of glycemic control in dogs with diabetes mellitus.
- The established reference interval aids in the clinical interpretation of canine HbA1c results.
Background:
Hemoglobin A1c (HbA1c) provides a reliable measure of glycemic control over 2-3 months in human diabetes mellitus. In dogs, presence of HbA1c has been demonstrated, but there are no validated commercial assays.
Objective:
The purpose of the study was to validate a commercially available automated immunoturbidimetric assay for canine HbA1c and determine an RI in a hospital population.
Methods:
The specificity of the assay was assessed by inducing glycosylation in vitro using isolated canine hemoglobin, repeatability by measuring canine samples 5 times in succession, long term inter-assay imprecision by measuring supplied control materials, stability using samples stored at 4°C over 5 days and -20°C over 8 weeks, linearity by mixing samples of known HbA1c in differing proportions, and the effect of anticoagulants with paired samples. An RI was determined using EDTA-anticoagulated blood samples from 60 nondiabetic hospitalized animals of various ages and breeds. Hemoglobin A1c was also measured in 10 diabetic dogs.
Results:
The concentration of HbA1c increased proportionally with glucose concentration in vitro. For repeat measurements, the CV was 4.08% (range 1.16-6.10%). Samples were stable for 5 days at 4°C. The assay was linear within the assessed range. Heparin- and EDTA-anticoagulated blood provided comparable results. The RI for HbA1c was 9-18.5 mmol/mol. There was no apparent effect of age or breed on HbA1c. In diabetic dogs, HbA1c ranged from 14 to 48 mmol/mol.
Conclusions:
The assay provides a reliable method for canine HbA1c measurement with good analytic performance.