Creatinine and cystatin C were measured in serum samples. Creatinine was measured using the enzymatic method (Roche Creatinine Plus ver.2 assay), which is standardized against the Isotope Dilution-Mass Spectrometry method. Cystatin C was measured using the immunoturbidimetric method (Tina-quant Cystatin C Gen. 2, Roche), which is standardized and traceable against ERM-DA471/IFCC reference material. Both tests were measured using the Roche cobas 8000 c 702 (Roche Diagnostics, Mannheim, Germany).
Tina quant cystatin c gen 2
Manufactured by Roche
Sourced in Switzerland, Germany
Tina-quant Cystatin C Gen. 2 is a laboratory equipment product manufactured by Roche. It is used for the quantitative determination of cystatin C in human serum and plasma samples.
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4 protocols using tina quant cystatin c gen 2
1
Serum Creatinine and Cystatin C Measurement
2
Cystatin C, Creatinine, and BUN Assays
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Serum cystatin C was measured by a turbidimetric immunoinhibition assay using the cystatin kit (Tina-quant cystatin C Gen. 2, Roche) with an automatic biochemical analyzer (Cobas c-system, Roche, Switzerland). This method is traceable to a primary reference material with values assigned by the international cystatin C reference material (ERM-DA471/IFCC) as a calibrator19 (link). This procedure had a total coefficient of variation of 2.2% at a cystatin C level of 1.0 mg/L and of 1.4% at a level of 4.0 mg/L. Reference range 0.55–1.09 mg/L. Serum creatinine was determined by a traceable method (enzymatic assay calibrated against the National Institute of Standards and Technology standard reference material [SRM 967]) with an automatic biochemical analyzer (Cobas c-system, Roche, Switzerland)20 (link). The total coefficient of variation is < 4.0%, and the reference range is 44–133 μmol/L. Serum Bun was determined by an enzymatic method with an automatic biochemical analyzer (Cobas c-system, Roche, Switzerland) according to the kit protocol. This method is traceable to a primary reference material (SRM 909b). This procedure had a total coefficient of variation of 1.2% at a level of 7.2 mmol/L and of 0.7% at a level of 35.1 mmol/L. The reference range is 2.9–8.2 mmol/L.
3
Cystatin C Measurement Protocol
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Serum cystatin C measurements were performed with the Tina-Quant Cystatin C Gen.2 assay (Roche) on cobas 8000 Modular Analyzer (Roche) by an external accredited laboratory (Labor Staber, Munich, Germany), using the human serum certified reference material (CRM) ERM-DA471/IFCC (European Commission Joint Research Centre, Institute for Reference Materials and Measurements, Belgium) as cystatin C calibrator [29 (link)].
4
Evaluation of GFR by MDRD and CKD-EPI
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Serum Cr was measured by using the Toshiba 200-FR analyzer (Toshiba Medical Systems, Tokyo, Japan) with the Roche calibrator and reagent (Roche Diagnostics, Indianapolis, IN, USA), which is traceable with the IDMS reference method. The dynamic measuring range was 0.2–25 mg/dL, and the mean within-laboratory imprecision was 1.35% during the study period. CysC was determined with the Roche Cobas 8000 modular system using Roche Tina-quant Cystatin C Gen 2, a particle enhanced immunonephelometric assay. The measurement range was 0.4–8.0 mg/L, and the mean within-laboratory imprecision was 2.7% during the study period. eGFRs were calculated by using the MDRD Study equation and CKD-EPI equations using Cr, CysC, or Cr-CysC as variables with demographic variables [9 (link)10 (link)11 (link)]. To assess eGFR, rather than using arbitrary categories, we adopted six eGFR categories according to the KDIGO guidelines: ≥90 mL/min/1.73 m2; 60–89 mL/min/1.73 m2; 45–59 mL/min/1.73 m2; 30–44 mL/min/1.73 m2; 15–29 mL/min/1.73 m2; <15 mL/min/1.73 m2 [19 ]. GFR ≥60 mL/min/1.73 m2 was considered normal.
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