S100B serum levels at admission and subsequent sampling were retrospectively reviewed. In most patients, samples were drawn on admission and daily afterwards. Some patients had several samples drawn on the same day (because of acute deterioration for instance), and only the first was used for analysis. In some cases, mostly after several days of normal S100B levels, sampling was discontinued until occurrence of neurological deterioration. Normal S100B level was arbitrarily defined as <0.1 μg/L in this study. A secondary increase of S100B serum level was defined as an absolute elevation of 0.1 μg/L at any point in time. Analysis was stopped at 30 days after admission. Physicians were aware of S100B serum level values in real time. Since December 2, 2010, S100B has been assayed on ModularE170 analyzer (Roche, Mannheim, Germany). Results assessed before that date were measured on a LiaisonXL (Diasorin, Saluggia, Italy); therefore, these values were corrected as (concentration − 0.01)/2.28, according to the linear regression between the two methods as previously reported [16 (link)].
Modular e170 analyzer
Manufactured by Roche
Sourced in Switzerland, Germany, France
The Roche Modular E170 Analyzer is an automated instrument used for in vitro diagnostic testing. It is designed to perform various immunoassay tests, including those for hormones, therapeutic drug monitoring, and other clinical chemistry analyses. The Modular E170 Analyzer offers high throughput and reliable performance to support clinical laboratories in their diagnostic procedures.
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Jouan centrifuge, by Thermo Fisher Scientific (2 mentions)
Cobas 8000 analyzer, by Roche (2 mentions)
Liaison xl, by DiaSorin (1 mentions)
Immulite 2000 immunoassay system, by Siemens (1 mentions)
Au5800 automatic analyzer, by Beckman Coulter (1 mentions)
Spss software, by IBM (1 mentions)
Elecsys assay, by Roche (1 mentions)
Hbsag qual 2 test, by Abbott (1 mentions)
Lightcycler 480, by Roche (1 mentions)
Coulter lh 750 hematology analyzer, by Beckman Coulter (1 mentions)
Architect i2000 analyzer, by Abbott (1 mentions)
Immage 800 analyzer, by Beckman Coulter (1 mentions)
Plasma low hb system, by HemoCue (1 mentions)
D p modular analyzer, by Roche (1 mentions)
Au5400 chemistry analyzer, by Olympus (1 mentions)
Cobas b 101 analyzer, by Roche (1 mentions)
Cobas c501 analyzer, by Roche (1 mentions)
Accu chek compact plus, by Roche (1 mentions)
Jouan mr23i, by Thermo Fisher Scientific (1 mentions)
25 protocols using modular e170 analyzer
1
Serum S100B Levels in Neurological Deterioration
2
Standardized Vitamin D Measurement
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Serum 25-hydroxyvitamin D (vitamin D) concentrations were measured through Cobas electrochemiluminescence immunoassays (ECLIA) on the Modular E170 analyzer (Roche Diagnostics, Mannheim, Germany).
This method was standardized against LC-MS/MS [43 (link)] which, in turn, was standardized against the NIST standard [44 (link)]. The performance of this assay was validated using Elecsys reagents, human sera and controls, performed according to the protocol EP5-A2 of the Clinical and Laboratory Standards Institute (CLSI).
The functional sensitivity of the assay was 4.01 ng/mL (CV 18.5%). The assay specificity (reflecting by the percentage of cross reactivity with other metabolites) was 100% for 25(OH)D3, 92% for 25(OH)D2, and 91% for the C3 epimer of 25(OH)D3.
A vitamin D level of 30 ng/mL or higher was considered adequate, whereas a concentration from 12 to 30 ng/mL was considered a vitamin D insufficiency and below 12 ng/mL a vitamin D deficiency.
This method was standardized against LC-MS/MS [43 (link)] which, in turn, was standardized against the NIST standard [44 (link)]. The performance of this assay was validated using Elecsys reagents, human sera and controls, performed according to the protocol EP5-A2 of the Clinical and Laboratory Standards Institute (CLSI).
The functional sensitivity of the assay was 4.01 ng/mL (CV 18.5%). The assay specificity (reflecting by the percentage of cross reactivity with other metabolites) was 100% for 25(OH)D3, 92% for 25(OH)D2, and 91% for the C3 epimer of 25(OH)D3.
A vitamin D level of 30 ng/mL or higher was considered adequate, whereas a concentration from 12 to 30 ng/mL was considered a vitamin D insufficiency and below 12 ng/mL a vitamin D deficiency.
3
Maternal Serum Biomarker Assay Protocol
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All maternal serum samples were assayed for b-hCG, progesterone, inhibin A, activin A and C-reactive protein (CRP) using commercial assays. Maternal serum progesterone and hCG assays were performed on a Modular E170 Analyzer (Roche Diagnostics, Vilvoorde, Belgium) with an electrochemiluminescence competitive methods. High sensitivity (hs) CRP assay was performed using an immunoturbidimetric method on a Modular P Analyzer (Roche Diagnostics, Vilvoorde, Belgium) with a quantitation limit of 0.5 mg/L. Maternal serum inhibin A and activin A assay was performed on a IMMULITE 2000 immunoassay system (Siemens, Brussels, Belgium) with an enzyme-labeled chemiluminescent immunometric method.
4
Hepatitis B Serological Markers Measurement
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With 3 mL of serum, both HBsAg and anti-HBs were measured in the same sample using Microparticle enzyme immunoassay (MEIA) in 2002-2008 and Electrochemiluninescence immunoassay (ELCIA) in 2008-2018 according to the instructions of the Modular E170 analyzer (Roche Diagnostics, Mannheim, Germany) which was consistently employed during the study period. The percent agreement for MEIA and ELCIA was 92.8% in a previous study (Whang and Shin, 2002) . This assay principle uses a two-step enzyme immunoassay sandwich method, combined with a final fluorescence detection step. For HBsAg serostatus, level of <1.0, and 1.0 (!1.0) IU/mL were regarded to be negative and positive, respectively. Anti-HBs !10 mIU/mL was recorded as anti-HBs seropositivity.
5
Immunogenicity Evaluation of HBsAg
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Frozen serum samples collected from the immunized mice were transferred to the reference lab at The First Affiliated Hospital of Wenzhou Medical University for assays of HBsAg and serum ALT levels. The Modular E170 analyzer (Roche Diagnostics, Basel, Switzerland) was employed for HBsAg assay. ALT activity was quantified by the AU5800 automatic analyzer (Beckman Coulter, Inc.).
Statistical analysis. Data analysis was performed using SPSS software (version 18.0; SPSS Inc., Chicago, IL, USA). Data are expressed as the mean ± standard deviation. Statistical analysis was performed using one-way analysis of variance followed by LSD-t or Dunnett's T3 post hoc tests for multi group comparisons. P<0.05 was considered to indicate a statistically significant difference.
Statistical analysis. Data analysis was performed using SPSS software (version 18.0; SPSS Inc., Chicago, IL, USA). Data are expressed as the mean ± standard deviation. Statistical analysis was performed using one-way analysis of variance followed by LSD-t or Dunnett's T3 post hoc tests for multi group comparisons. P<0.05 was considered to indicate a statistically significant difference.
6
Thyroid and Vitamin D Biomarker Assessment
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Serum fT4 and TSH were measured using a fT4-CTX radioimmunoassay (RIA) kit (Diasorin, Saluggia, Italy) and RIA-gnost ® hTSH kit (Cisbio Bioassays, Codolet, France), respectively. Serum TSHR-Ab was determined using the TRAK human RIA (BRAHMS) kit (Thermo Scientific, Henningsdorf, Berlin, Germany). TPO-Ab and Tg-Ab were measured using the TM Ab immunoradiometric assay (IRMA) CT kit (RADIM, Pomezia, Italy) and Tg-Ab using the one step CT RIA kit (RADIM). Serum concentrations of 25(OH)D were measured using a Roche Modular E170 Analyzer and an electrochemiluminescence immunoassay (ECLIA; Roche Diagnostics ® , Penzberg, Germany). Serum PTH levels were assessed using a total intact PTH IRMA kit (IBL international GMBH, Hamburg, Germany). Serum calcium, albumin, and phosphorus were measured using a model TBA-200FR instrument (Toshiba Medical System, Tokyo, Japan). Corrected calcium levels were calculated on the basis of albumin levels. Vitamin D insufficiency and deficiency were defined as serum 25(OH)D levels <75 nmol/L (30 ng/mL) and <50 nmol/L (20 ng/mL), respectively.
7
Quantification of HBsAg and HBeAg
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For HBsAg-positive samples, qHBsAg was quantified using the Elecsys assay with the Modular E170 analyzer (Roche Diagnostics, Meylan, France). Similarly for HBeAg-positive samples, qHBeAg was quantified using the Elecsys assay. This automated electrochemiluminescence immunoassay gives qHBeAg levels in a semi-quantitative manner and is interpreted as the ratio of sample relative light units to a cut-off value. These units were converted to Paul Ehrlich Institute units (PEIU/mL) from a previously established protocol. 17
8
Quantifying HBV Biomarkers in Chronic Infection
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Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were quantified at baseline. From frozen samples stored at -80°C, HBV-DNA viral loads were quantified at baseline and every follow-up visit using an in-house PCR-based assay (Light Cycler 480, Roche, Boulogne-Billancourt, France) with a detection limit of 12 copies/mL. 16 HBsAg was detected using the HBsAg Qual II test (Architect, Abbott Laboratories, Rungis, France) at baseline and during follow-up. HBe antigen (HBeAg) and anti-HBe antibodies (anti-HBeAb) were detected using the Elecsys assay (Roche Diagnostics, Meylan, France) for the same visits. For HBsAg-positive samples, qHBsAg was quantified using the Elecsys assay with Modular E170 analyzer (Roche Diagnostics, Meylan, France). HBsAg-seroclearance was defined as loss of HBsAg from the previous visit.
9
Hematological Changes in Endurance Athletes
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Using sterile techniques, blood (20 mL) was drawn from an antecubital vein from each subject 1 week before and immediately after the race. All specimens were refrigerated and transported to the laboratory within 4 h of sampling. Red blood cell (RBC) count, hemoglobin, hematocrit, mean cell volume (MCV), mean cell hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width, coefficient of variation (RDW-CV) and reticulocyte were performed on a Coulter LH 750 Hematology Analyzer (Beckman Coulter, Miami, FL, USA), which was based on impedance detection for counting and sizing the blood cells. The evaluated plasma haptoglobin was assayed by the rate nephelometry on an Immage 800 Analyzer (Beckman Coulter, Fullerton, CA, USA). Free plasma hemoglobin was tested with a HemoCue Plasma/Low Hb System (HemoCue, Lake Forest, CA, USA) utilizing the azide-methemoglobin method. Ferritin levels were determined by an Architect I-2000 Analyzer (Abbott Diagnostics, Abbott Park, IL, USA), which used a chemiluminescent microparticle immunoassay. Iron and unsaturated iron-binding capacity (UIBC) were measured on a Modular E170 Analyzer (Roche Diagnostics, Mannheim, Germany) using an electrochemiluminescence immunoassay.
10
Nutritional Assessment of Hemodialysis Patients
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Patient demographic data, concurrent CV disease history, waist circumference (WC), and smoking status were recorded. Venous blood was sampled after fasting for more than 8 hours before the HD patient’s mid-week HD session. Whole blood samples were obtained for haemoglobin measurement, whereas serum was used for determining creatinine, calcium, phosphorus, potassium, uric acid, albumin, and lipid profiles. Intact parathyroid hormone levels were determined using immunoassay (Roche Modular E170 Analyzer, Roche Diagnostics GmbH, Mannheim, Germany). hs-CRP levels were determined using an immunonephelometric method with a Tina-quant CRP (Latex) ultrasensitive assay (D & P Modular Analyzer, Roche Diagnostics GmbH). Estimated GFRs were measured using the Modification of Diet in Renal Disease Study equation as follows: GFR (in mL/min/1.73 m2) = 175 × (Scr, serum creatinine, in mg/dL)−1.154 × (Age, in years)−0.203 × (0.742, if female).
The nutritional status of the participants was quantified using the GNRI, calculated on the basis of serum albumin levels and body weight as follows: GNRI = [14.89 × albumin level (in g/dL)] + [41.7 × body weight/WLo], where WLo is the ideal body weight calculated in the Lorentz equation. The GNRI has been validated in HD patients, and a higher GNRI indicates high nutritional status32 (link).
The nutritional status of the participants was quantified using the GNRI, calculated on the basis of serum albumin levels and body weight as follows: GNRI = [14.89 × albumin level (in g/dL)] + [41.7 × body weight/WLo], where WLo is the ideal body weight calculated in the Lorentz equation. The GNRI has been validated in HD patients, and a higher GNRI indicates high nutritional status32 (link).
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