Elecsys hbsag 2 quant

Manufactured by Roche

Sourced in Germany, Switzerland, United States

The Elecsys HBsAg II Quant is a laboratory equipment product designed for the quantitative determination of hepatitis B surface antigen (HBsAg) in human serum and plasma samples. It provides a standardized and automated solution for the detection and measurement of HBsAg levels.

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7 protocols using elecsys hbsag 2 quant

Quantitative Hepatitis B Viral Assessment

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Whole‐blood samples were processed for serum and samples stored at –80°C until transfer to the bioanalytical laboratory. Quantitative HBV serology parameters assessed were HBV DNA (Cobas AmpliPrep/Cobas TaqMan, v2.0; Roche Diagnostics), HBsAg (Elecsys HBsAg II quant; Roche Diagnostics), HBeAg (Liaison; DiaSorin; E‐pos patients only). Qualitative assessments also included the presence of antibody to HBsAg and antibody to HBeAg (E‐pos patients only) as well as HBV genotyping and sequencing of archival samples, where available.

Yuen M., Schiefke I., Yoon J., Ahn S.H., Heo J., Kim J.H., Lik Yuen Chan H., Yoon K.T., Klinker H., Manns M., Petersen J., Schluep T., Hamilton J., Given B.D., Ferrari C., Lai C., Locarnini S.A, & Gish R.G. (2020). RNA Interference Therapy With ARC‐520 Results in Prolonged Hepatitis B Surface Antigen Response in Patients With Chronic Hepatitis B Infection. Hepatology (Baltimore, Md.), 72(1), 19-31.

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Serological HBV Markers and Fibrosis Indices

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Serological testing for HBV DNA, quantitative HBsAg, HBeAg, and HBV genotype were performed in a central laboratory. The following assays were used: HBV DNA by Roche real-time polymerase chain reaction assay (COBAS Ampliprep/COBAS Taqman HBV Test, Branchburg, NJ) with a lower limit of quantitation of 20 IU/mL; quantitative HBsAg by Elecsys HBsAg II Quant (Roche Diagnostics) with a lower limit of 0.05 IU/mL; qualitative HBeAg by EIA (Diasorin, ETI-EBK PLUS) and quantitative HBeAg by Elecsys HBeAg Test (Roche Diagnostics) with a lower limit of 0.3 IU/mL; and HBV genotype by mass spectrometry^{17 (link)}. When results from the central laboratory were not available, results from local laboratories at clinical sites were used for analysis.
Hyperlipidemia and diabetes were self-reported clinical diagnoses at entry into study. Fibrosis-4 index was calculated using the following equation: age [years] × AST [IU/L]/platelet count [expressed as platelets × 10⁹/L] × (ALT½[IU/L])^{18 (link)}. AST-to-platelet ratio index (APRI) was calculated as: (AST/upper limit of normal)/platelet count (expressed as platelets × 10⁹/L) × 100^{19 (link)}.

Zhou K., Wahed A.S., Cooper S., Di Bisceglie A.M., Fontana R.J., Ghany M.G., Khalili M., Lok A.S., Perrillo R., Lee W., Lau D., Sterling R., Janssen H.L, & Terrault N.A. (2019). Phase Transition is Infrequent Among North American Adults with e-Antigen Negative Chronic Hepatitis B and Low-level Viremia. The American journal of gastroenterology, 114(11), 1753-1763.

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Comprehensive Viral Hepatitis Screening

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ALT and viral markers (HBsAg, antibodies to HBsAg [anti‐HBs], HBeAg, anti‐HBe, hepatitis C and D virus antibodies, anti‐HIV) were determined using conventional serological assays. Serum samples at baseline were stored at −20°C. Up to 2002, serum HBV DNA levels were analysed on stored serum samples with the branched DNA signal amplification assay (Chiron Diagnostics, Emeryville, CA) lower limit of detection of 0.7 mEq/mL (7.00 × 10⁵ IU/mL). Thereafter, HBV DNA quantification was performed by ABI Prism 7900HT (Applied Biosystems, Thermo Fisher Scientific, Waltham, MA) with a detection limit of 100 IU/mL and from 2015 on by a commercial PCR assay (Abbott RealTime HBV assay; Abbott Molecular Inc, Des Plaines, IL) with a sensitivity of 10 IU/mL. Serum qHBsAg was measured on the Elecsys HBsAg II quant (Roche Diagnostics, Penzberg, Germany) or Architect HBsAg QT (Abbott Diagnostics, IL) assay.

Koc Ö.M., Robaeys G., Topal H., Bielen R., Busschots D., Fevery J., Koek G.H, & Nevens F. (2020). Outcome in Caucasian patients with hepatitis B e antigen negative chronic infection: A long‐term observational cohort study. Journal of Medical Virology, 92(12), 3373-3380.

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Serological Characterization of HBV Infection

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Serological testing for markers of HBV infection was done centrally, as previously described.^{15 (link)} Briefly, HBV-DNA levels were determined using a real-time PCR assay (COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0; Roche Molecular Diagnostics, Branchburg, NJ). HBV genotype was determined by mass spectrometry at the Molecular Epidemiology and Bioinformatics Laboratory in the Division of Viral Hepatitis at the Centers for Disease Control and Prevention (CDC) with mass spectrometry,^{16 (link)} while quantitative hepatitis B e antigen (HBeAg) and HBsAg levels were measured by Elecsys HBeAg II Quant and Elecsys HBsAg II Quant assays, respectively (Roche Molecular Systems, Inc).^{17 (link),18} The lowest detectable value for HBV-DNA was 10 IU/mL, for HBeAg was 0.3 IU/mL, and for HBsAg was 0.05 IU/mL; the lowest quantifiable value for HBV-DNA was 20 IU/mL. If not available from the central laboratory, local data were used. Serum biochemical testing including serum aminotransferase and bilirubin levels was performed at local laboratories.

Lieske J.C., Swift H., Martin T., Patterson B, & Toback F.G. (1994). Renal epithelial cells rapidly bind and internalize calcium oxalate monohydrate crystals.. Proceedings of the National Academy of Sciences of the United States of America, 91(15), 6987-6991.

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Quantification of HBsAg and HBV-DNA

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Cell supernatants were collected, clarified by centrifugation at 4000 g for 5 min and used to quantify HBsAg by Elecsys^® HBsAg II quant (Roche, Basel, Switzerland) and HBV-DNA with cobas^® HBV Test (Roche, Basel, Switzerland).

Salpini R., Piermatteo L., Battisti A., Colagrossi L., Aragri M., Yu La Rosa K., Bertoli A., Saccomandi P., Lichtner M., Marignani M., Maylin S., Delaugerre C., Morisco F., Coppola N., Marrone A., Iapadre N., Cerva C., Aquaro S., Angelico M., Sarmati L., Andreoni M., Verheyen J., Ceccherini-Silberstein F., Levrero M., Perno C.F., Belloni L, & Svicher V. (2020). A Hyper-Glycosylation of HBV Surface Antigen Correlates with HBsAg-Negativity at Immunosuppression-Driven HBV Reactivation in Vivo and Hinders HBsAg Recognition In Vitro. Viruses, 12(2), 251.

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Evaluating HBsAg Secretion and Antigenicity

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In order to better unravel the role of the identified N-linked glycosylation sites in affecting HBsAg secretion or antigenic properties, a plasmid encoding the HBsAg linked to a streptavidin-tag (version II, IBA Lifesciences, Göttingen, Germany) (strep-tag) was used to transfect the Huh7 cells. The strep-tag is upstream the gene encoding HBsAg, thus resulting at the N-terminus of the corresponding protein.
After 72 h post transfection, cell supernatants were collected, clarified by centrifugation at 4000 g for 5 min, and used for the quantification of strep-tag HBsAg. In particular, the amount of strep-tagged HBsAg released in culture supernatants was quantified by using two assays targeting the major hydrophilic region of HBsAg (Monolisa™ HBs Ag ULTRA [Bio-Rad Laboratories, Hercules, CA, USA] and Elecsys^® HBsAg II quant [Roche, Basel, Switzerland]) and by using a specifically-designed ELISA capable to recognize the Strep-tag portion linked to the HBsAg (defined hereafter as Strep-tag ELISA). Differently from the commonly used HBsAg assays, the Strep-tag based ELISA is not influenced by HBsAg modifications, giving the advantage to discriminate between a reduction of HBsAg recognition (due to an altered HBsAg antigenic properties) and a decrease in HBsAg secretion.
For each mutant, at least 3 independent transfection experiments were performed, each carried out in duplicate.

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Quantitative Assessment of HBV Markers

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Serological testing for markers of HBV infection was done centrally, as previously described. 15 Briefly, HBV-DNA levels were determined using a real-time PCR assay (COBAS AmpliPrep/COBAS TaqMan HBV Test, v2.0; Roche Molecular Diagnostics, Branchburg, NJ). HBV genotype was determined by mass spectrometry at the Molecular Epidemiology and Bioinformatics Laboratory in the Division of Viral Hepatitis at the Centers for Disease Control and Prevention (CDC) with mass spectrometry, 16 while quantitative hepatitis B e antigen (HBeAg) and HBsAg levels were measured by Elecsys HBeAg II Quant and Elecsys HBsAg II Quant assays, respectively (Roche Molecular Systems, Inc). 17, 18 The lowest detectable value for HBV-DNA was 10 IU/mL, for HBeAg was 0.3 IU/mL, and for HBsAg was 0.05 IU/mL; the lowest quantifiable value for HBV-DNA was 20 IU/mL. If not available from the central laboratory, local data were used. Serum biochemical testing including serum aminotransferase and bilirubin levels was performed at local laboratories.

Kleiner D.E., Lisker-Melman M., Wahed A.S., Bhan A.K., Nalesnik M.A., Choi E.K., Leonard K.K., Ghany M.G., Chung R.T, & Di Bisceglie A.M. (2023). Immunostaining for hepatitis B viral antigens in liver: Association with clinical, biochemical, and virologic features of disease. Journal of gastroenterology and hepatology, 38(6).

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