Urisys 1100

The blood samples were analyzed for glucose using contour sticks and device (Bayer Consumer Care AG, Basel, Switzerland). The urine samples were tested for pH and ketones using Combur^_7Test sticks read in an Urisys 1100 (Roche Diagnostics GmbH, Germany). The urine was also tested for protein, leukocytes and erythrocytes/hemoglobin content immediately after sampling with Combur^_7Test sticks read in Urisys 1100 (Roche).

At the time of diagnosis, we measured fasting C-peptide and HbA_1c, anti-glutamic acid decarboxylase (GAD), anti-insulin and anti-islet antigen 2 (IA-2) autoantibodies, thyroid-stimulating hormone (TSH), and urine ketones. We also recorded the presence of acidosis episodes and the need for insulin treatment three to six months after the diagnosis and performed abdominal ultrasonography. We repeated the HbA_1c measurements six and 12 months after the diagnosis. We determined HbA_1c by high-performance liquid chromatography using the Diabetes Control and Complications Trial (DCCT) % units [35 (link)]. We measured the fasting serum C-peptide and TSH levels by chemiluminescent immunoassay method using Cobas E 411 analyzer (Roche diagnostics, Basel, Switzerland). We detected the autoantibodies using radioimmunoassays from DIAsource ImmunoAssays S.A. (Louvain-la-neuve, Belgium). Furthermore, we measured ketone bodies using Combur-Test strip with the Urisys 1100 semi-automated urine testing analyzer (Roche Diagnostics, Basel, Switzerland).

Routine biochemical urine tests were performed using Cobas Urisys 1100 analyser (Roche Cobas, Swiss) using appropriate test strips. Energy production (citric acid cycle intermediates—citrate, cis-aconitate, isocitrate, alfa-ketoglutarate, succinate, fumarate, malate, and hydroxymethylglutarate), neurotransmitter metabolism (quinolinate, picolinate, vanilmandelate, homovanillate, 5-hydroxyindolacetate, and kynurenate) biomarkers, and porphyrin content (uroporphyrins, heptacarboxylporphyrins, hexacarboxylporphyrins, pentacarboxylporphyrins, coproporphyrin I concentration, coproporphyrin III, total porphyrins, precoproporphyrin I, precoproporphyrin II, precoproporphyrin III concentration, and total precoproporphyrins) in the urine were performed using LC/MS-MS spectrophotometry (Organix Comprehensive Profile; Metametrix, Inc, Duluth, GA). Urine samples were collected in accordance with instructions provided by Metametrix Clinical Laboratory. In brief, participants were instructed to void (empty bladder) before going to bed and then place a collection basin over their home toilets. Participants used a pipette to place 12 mL of their first morning sample plus any overnight sample into a test tube. At the laboratory, urine samples were frozen and then shipped to Metametrix Clinical Laboratory for biochemical analyses.