All biochemical parameters (LDL-cholesterol, HDL-cholesterol, triglycerides, glucose) were determined within 1 week of cardiovascular examination by standard methods on a Roche-Modular Autoanalyzer (Milan, Italy). Glycosylated hemoglobin (HbA1c) was measured by high-performance liquid chromatography and standardized against DCCT standard.
Modular autoanalyzer
Manufactured by Roche
Sourced in Italy, United States, Japan, Germany
The Roche-Modular Autoanalyzer is a laboratory instrument designed for automated chemical analysis. It is capable of performing a variety of clinical chemistry tests on samples, such as determination of analyte concentrations in biological fluids. The Roche-Modular Autoanalyzer is a modular system that can be configured to meet the specific needs of a laboratory.
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Lab products found in correlation
Proseek multiplex cvd96 96 reagents kit, by Olink (1 mentions)
Elx800tm, by Agilent Technologies (1 mentions)
Rat nf κb elisa kit, by Elabscience (1 mentions)
Cobas integra 800, by Roche (1 mentions)
Glucose analyzer 2, by Beckman Coulter (1 mentions)
Immage, by Beckman Coulter (1 mentions)
Quantikine, by R&D Systems (1 mentions)
Rx daytona chemistry analyzer, by Randox (1 mentions)
10 protocols using modular autoanalyzer
1
Cardiovascular Biomarker Determination
2
Cardiovascular Biomarkers Analysis Protocol
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Plasma levels of low density lipoprotein cholesterol (LDL), HDL, TG and glucose were determined within 1 week of CV examination by standard methods on a Roche-Modular Autoanalyzer (Milan, Italy). Glycosylated hemoglobin (HbA1c) was measured by high-performance liquid chromatography and standardized against Diabetes Control and Complications Trial (DCCT ) standard. Plasma levels of biomarkers reflecting inflammation (IL-6 and IL-18) and extracellular matrix proteolysis (MMP-12) were analyzed by the Proximity Extension Assay (PEA) technique using the Proseek Multiplex CVD96×96 reagents kit (Olink Bioscience, Uppsala, Sweden) at the Clinical Biomarkers Facility, Science for Life Laboratory, Uppsala as previously described [9 (link), 26 (link)]. All samples were analyzed in the same run. Data analysis was performed by a preprocessing normalization procedure using Olink Wizard for GenEx (Multid Analyses, Sweden). Values are presented as arbitrary units (AU). Data regarding intra- and inter-assays variations as well as general calibrator curves to calculate the approximate concentrations are available on the OLINK homepage (http://www.olink.com ).
3
GWAS of Lipid Traits in Prediabetes
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To identify genetic loci associated with lipid traits, including TG, TC, HDL-C, and LDL-C, in the context of prediabetes among Southern Han Chinese, we conducted a GWAS (Figure 1
This study was approved by the Ethical Committee of the Hainan Affiliated Hospital of Hainan Medical University (Med-Eth-Re (2019) 18) and adhered to ethical standards set forth by the committee and the Declaration of Helsinki. All participants provided written informed consent after being fully informed of the study’s purpose.
This study was approved by the Ethical Committee of the Hainan Affiliated Hospital of Hainan Medical University (Med-Eth-Re (2019) 18) and adhered to ethical standards set forth by the committee and the Declaration of Helsinki. All participants provided written informed consent after being fully informed of the study’s purpose.
4
Serum Biomarker Measurement Protocol
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Collected blood samples were centrifuged at 2000 rpm for 15 min (with the use of re-frigerated centrifuge) to obtain serum samples. Thereafter, liver enzymes (ALT, AST and ALP), urea and creatinine levels were measured colorimetrically utilizing the methods used by routine Roche modular autoanalyzer equipment (Roche modular autoanalyzer, Tokyo, Japan) [28 (link)].
5
Serum Biomarker Levels in Migraine
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Blood samples were taken from the control group and from the migraine group. Among the migraine patients, this was done both when they were experiencing migraine headaches and when they were not. The venous blood samples were instantly centrifuged at 3000 rpm for 10 min at 4 °C and then poured into Eppendorf tubes. The serum samples were transferred on ice and stored at -80 °C for three months until the end of the study period.
The serum vaspin, visfatin, chemerin and IL-18 levels were measured using commercially available enzyme-linked immunosorbent assay kits (YL Biont, China). The absorbance was read at 450 nm and recorded using an absorbance microtiter plate reader (ELx800 TM ; BioTek Instruments, USA). The serum glucose, total protein, albumin, globulin, cholesterol and triglyceride levels were determined using routine colorimetric methods in an autoanalyzer (Roche Modular Autoanalyzer; Roche, Tokyo, Japan).
The serum vaspin, visfatin, chemerin and IL-18 levels were measured using commercially available enzyme-linked immunosorbent assay kits (YL Biont, China). The absorbance was read at 450 nm and recorded using an absorbance microtiter plate reader (ELx800 TM ; BioTek Instruments, USA). The serum glucose, total protein, albumin, globulin, cholesterol and triglyceride levels were determined using routine colorimetric methods in an autoanalyzer (Roche Modular Autoanalyzer; Roche, Tokyo, Japan).
6
Plasma Lipid Measurement Protocol
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We generated plasma using Microtainer plasma separator tubes (Becton Dickinson). Veterinary Diagnostic Services (Marshfield Laboratories) and measured the levels of cholesterol, β-hydroxybutyrate, FFA, and triglycerides by an automated analyzer (Roche Modular Autoanalyzer). We assayed liver triglycerides using the triglyceride GPO reagent as previously described [8 (link)].
7
Comprehensive Biomarker Assessment Protocol
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The concentrations of glucose, urea, uric acid, creatinine, and alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transpeptidase (GGT) activities were determined by the modular autoanalyzer (Roche, Germany). The NF-κB levels were measured in the obtained serum samples using the commercial Rat NF-κB ELISA Kit (Catalog Number: E-EL-R0673, Elabscience, China). The serum 8-OHdG (oxidative DNA damage) level was determined using a commercial kit (Enzo Life Science Company of DNA Damage) ELISA kit (Catalog Number: ADI-EKS-350). The levels of glycosylated hemoglobin (HbA1c) were analyzed in whole blood using a commercial kit (Roche, Germany) and an autoanalyzer (Roche Cobas Integra 800) on the same day.
8
Fasting Blood Biomarkers Evaluation
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Blood samples were drawn after an overnight fast (10–12 h) for plasma glucose, determined by the glucose oxidase method (Glucose Analyzer 2, Beckman Instruments, Fullerton, CA, USA), serum insulin, determined by a Microparticle Enzyme Immunoassay (MEIA) (AxSym Insulin Kit, Abbot, IL, USA), TC, HDL-C, and TG, analyzed using the Roche Modular auto analyzer and enzymatic colorimetric assays, and LDL-C calculated using the Friedewald formula [27 (link)]. Homeostatic model assessment (HOMA-IR) was estimated from fasting insulin and glucose levels as previously described [28 (link)]. Serum concentrations of hsCRP (Immun Diagnostik AG, Bensheim, Germany) were analyzed using commercially available ELISA kits according to the manufacturer’s protocols.
9
Antiretroviral Therapy Metabolic Effects
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As ART therapy can affect VD levels and subclinical atherosclerosis, patients were classified into three groups: (1) those who were naïve to ART; (2) the non-nucleoside group: EFV or nevirapine plus two or three NRTIs; and (3) the PI group: PI plus two or three NRTIs.
Lipid and carbohydrate metabolism variables: fasting glycaemia and basal lipid profile were measured; impaired fasting glucose (IFG) was defined as 5.5 to 7 mmol/L in at least two measurements; glucose was measured using the hexokinase method (Modular auto-analyzer; Roche Diagnostics®, Grenzach, Germany), and lipid profile was measured by calorimetric enzymatic techniques (Modular auto-analyzer).
Systemic inflammatory markers: we measured high-sensitivity C-reactive protein (hsCRP) (turbidimetry kinetics; IMMAGE, Beckmann Coulter, Inc., Chasca, MN, USA), plasminogen activator inhibitor-1 (PAI-I), tumour necrosis factor-alpha (TNF-α), soluble forms of TNF-1 and -2 receptors and interleukin-6 (IL-6) (enzyme immunoassay; Quantikine, R&D Systems, Abingdon, UK). Lipodystrophy was determined using a standard questionnaire based on physical examination [15 (link)].
Lipid and carbohydrate metabolism variables: fasting glycaemia and basal lipid profile were measured; impaired fasting glucose (IFG) was defined as 5.5 to 7 mmol/L in at least two measurements; glucose was measured using the hexokinase method (Modular auto-analyzer; Roche Diagnostics®, Grenzach, Germany), and lipid profile was measured by calorimetric enzymatic techniques (Modular auto-analyzer).
Systemic inflammatory markers: we measured high-sensitivity C-reactive protein (hsCRP) (turbidimetry kinetics; IMMAGE, Beckmann Coulter, Inc., Chasca, MN, USA), plasminogen activator inhibitor-1 (PAI-I), tumour necrosis factor-alpha (TNF-α), soluble forms of TNF-1 and -2 receptors and interleukin-6 (IL-6) (enzyme immunoassay; Quantikine, R&D Systems, Abingdon, UK). Lipodystrophy was determined using a standard questionnaire based on physical examination [15 (link)].
10
Biochemical Assays for Metabolic Markers
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Plasma glucose was measured using the hexokinase method on a Randox RX Daytona Chemistry Analyzer. Enzymatic colorimetric assays were used to measure triglycerides, total cholesterol and high-density lipoprotein cholesterol using the Roche Modular Auto Analyzer, while low-density lipoprotein cholesterol was calculated using direct methods. Coefficients of variation calculated from running 40 separate samples in duplicate were 3.0% for glucose, 3.1% for cholesterol, 3.1% for triglycerides and 3.4% for insulin.
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