For diabetes patients, blood glucose was measured at diagnosis with a Selectra Pro S ELITech Group (Dieren, Netherlands). HbA1c was collected on the day of study assessment, and assayed using a BioRad D-10 analyser (Biorad Laboratories Inc., Hercules, USA). Fasting C-peptide and autoantibodies against glutamic acid decarboxylase 65 (GAD65) and islet antigen 2 (IA-2) were measured from frozen samples by commercially available ELISA kits (IBL, Hamburg, Germany) in Pakistan. Fasting C-peptide was measured in ng/mL. GAD65 and IA-2 autoantibodies were considered positive if levels were ≥30 IU/mL based on the standard curve, according to manufacturer’s recommendation. Similar ELISA formats have been challenged in Islet Autoantibody Standardization Programs with comparable sensitivity and specificity to radioimmunoassays (12 (link)).
D 10 analyser
Manufactured by Bio-Rad
Sourced in United States
The D-10 analyser is a fully automated high-performance liquid chromatography (HPLC) system designed for the quantitative measurement of hemoglobin variants and glycated hemoglobin (HbA1c) levels in human blood samples. The system performs sample preparation, separation, detection, and analysis in a single, integrated platform.
Automatically generated - may contain errors
3 protocols using d 10 analyser
1
Diabetes Biomarkers and Autoantibodies
2
Comprehensive Assessment of Type 2 Diabetes
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We used standardized questionnaires to record demographic and clinical information about duration of T2DM, years of formal education, health and medical history including vascular disease and risk factors, smoking, medication use, and alcohol use (g/day). We measured weight, height, and waist and hip circumferences and calculated Body Mass Index (BMI) as weight (kg)/height2 (m2); habitual physical activity by calculating mean number of steps per day using a Yamax pedometer worn over a seven-day period; mood using the 15-item Geriatric Depression Scale (GDS) [31 (link)]; and blood pressure (BP) using an Omron M4 sphygmomanometer in a sitting position as an average of 3 recordings from the right arm. Fasting plasma glucose was recorded using a Roche Cobas 6000 analyser with hexokinase determination and HbA1c using a Bio-Rad D10 analyser.
3
Biochemical Profiling for Adrenal Disorders
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Serum DHEA-S, cortisol and plasma ACTH were analysed using principles of electrochemiluminescence immunoassay by Cobas e411 (Roche Diagnostics, Germany). Serum was assessed for routine chemistry through the fully automated chemical analyser (Cobas c311 [Roche Diagnostics, Germany]). Plasma aldosterone, renin, normetanephrine, metanephrine and serum 17-hydroxyprogesterone (17-OHP) for those with elevated DHEA-S for the diagnosis of congenital adrenal hyperplasia (CAH) were analysed using enzyme-linked immunosorbent assay (ELISA) by the (DRG) R ELISA kit system (Germany). The diagnosis of primary aldosteronism was considered by measuring the aldosterone/renin ratio (ARR) >5.7.[ 17 ] The elevation of plasma metanephrine and/or normetanephrine four times above the normal value was used to establish the diagnosis of PCC.[ 18 ] HbA1c was measured by fully automated high-performance liquid chromatography system (Bio-Rad D-10 analyser). The diagnosis of diabetes mellitus was made according to the American Diabetic Association diagnostic criteria[ 19 ] or the patient is known to have diabetes and on treatment.
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