Realtime hcv

Manufactured by Abbott

Sourced in United States, Germany

The RealTime HCV is a laboratory equipment product designed for the detection and quantification of hepatitis C virus (HCV) RNA in human plasma or serum samples. It utilizes real-time PCR technology to provide accurate and reliable results for HCV viral load measurements.

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Close spelling variants of this product

Abbott RealTime HCV kit RealTime HCV quantitative assay RealTime HCV Amplification Reagent Kit RealTime HCV Viral Load Assay Abbott RealTime HCV GT II RealTime HCV Genotype II RealTime HCV assay

27 protocols using realtime hcv

Hepatitis E Virus Detection Protocol

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We designed 3 in-house–developed reverse transcription PCRs (RT-PCRs) to detect HEV (Technical Appendix Table 1). Hepatitis A virus (HAV) RNA and CMV DNA detections were performed using in-house nucleic acid amplification tests. HBV and HCV viral loads were quantified using commercial kits (COBAS TaqMan, Roche, Basel, Switzerland; and RealTime HCV, Abbott, Chicago, IL, USA, respectively).
We sequenced the PCR product of the pan-Orthohepevirus RT-PCR using the RT-PCR primers. Because the RNA-dependent RNA polymerase sequences of patient HEV isolates clustered with rat HEV-C strains, primers for complete genome amplification were designed by multiple alignment of rat HEV-C genomes in GenBank (Technical Appendix Table 2). We used these primers for complete genome sequencing of HEV in patient feces (strain LCK-3110). We constructed phylogenetic trees using MEGA6 with the general time reversible plus gamma model (12 (link)).

Sridhar S., Yip C.C., Wu S., Cai J., Zhang A.J., Leung K.H., Chung T.W., Chan J.F., Chan W.M., Teng J.L., Au-Yeung R.K., Cheng V.C., Chen H., Lau S.K., Woo P.C., Xia N.S., Lo C.M, & Yuen K.Y. (2018). Rat Hepatitis E Virus as Cause of Persistent Hepatitis after Liver Transplant. Emerging Infectious Diseases, 24(12), 2241-2250.

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HCV RNA Quantification in Plasma

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HCV RNA was quantified in plasma samples by Abbott Real Time HCV quantitative assay (Abbott, Wiesbaden, Germany) on Automated m2000sp/m2000rt platform, as per the manufacturer’s instructions. The lower limit of detection (LOD) and lower limit of quantification (LLOQ) of this assay is similar, ie, 12 IU/mL, and linear range of the assay is from 12 to 10⁸IU/mL.

Gupta E., Choudhary M.C., Upadhyay N., Singh G., Nayak S.L., Kumar M, & Sarin S.K. (2019). Lower Rates Of Naturally Occurring Resistance-Associated Substitutions (RASs) In Hepatitis C Virus (HCV)-Infected Chronic Kidney Disease (CKD) Patients Than In HCV-Infected Patients With Only Liver Disease. Infection and Drug Resistance, 12, 3635-3640.

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Evaluating SVR12 Rates in HCV Mono- and Co-infection

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Treatment effectiveness was measured by the percentage of patients achieving SVR12, defined as an HCV RNA below the lower limit of quantification or detection 12 weeks (70 to 126 days) after treatment completion. HCV RNA levels were determined using real-time polymerase chain reaction based on either the Roche COBAS TaqMan HCV or the Abbott RealTime HCV polymerase chain reaction assays. Patients were considered responders if HCV RNA was undetectable (lower limit of quantification or detection ≤50 IU/mL) or unquantifiable (lower limit of quantification ≤50 IU/mL). Virologic response was also assessed at the end of treatment (EoT).
To analyze the difference in SVR12 rates between patients with monoinfection and coinfection, both rates were compared assuming a non-inferiority margin of 5% (based on scientific discussion with researchers and clinicians) with a 90% confidence interval (CI) (Farrington-Manning method).

Manuel Sousa J., Vergara M., Pulido F., Sánchez Antolín G., Hijona L., Carnicer F., Rincón D., Salmerón J., Mateos-Muñoz B., Jou A., Polo-Lorduy B., Rubín Á., Escarda A., Aguilar P., Aldámiz-Echevarría T., García-Buey L., Carrión J.A., Hernández-Guerra M., Chimeno-Hernández S., Espinosa N., Morillas R.M., Andrade R.J., Delgado M., Gallego A., Magaz M., Moreno-Planas J.M., Estébanez Á., Rico M., Menéndez F., Sampedro B., Morano L., Izquierdo S., Zozaya J.M., Rodríguez M., Morán-Sánchez S., Lorente S., Martín-Granizo I., Von-Wichmann M.Á., Delgado M, & Manzanares A. (2019). Real-world evidence of the effectiveness of ombitasvir-paritaprevir/r ± dasabuvir ± ribavirin in patients monoinfected with chronic hepatitis C or coinfected with human immunodeficiency virus-1 in Spain. PLoS ONE, 14(11), e0225061.

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Evaluating HCV Treatment Outcomes

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SVR was defined by either an undetectable qualitative polymerase chain reaction (e.g. Amplicor HCV Test v2.0) or quantitative (e.g. Abbott Real Time HCV) HCV viral load below the assay's lower limit of detection, obtained at least 12 weeks after the end of HCV treatment. Estimated glomerular filtration rates (eGFR) were calculated using the serum creatinine-based Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation [27 (link)]. Alcohol consumption and illicit (injection and noninjection) cocaine and opiate substance use was obtained by self-report at each visit. Hazardous alcohol consumption was assessed using the Alcohol Use Disorders Identification Test (AUDIT-C) score [28 (link)]. Duration of HCV infection was estimated as the earliest date of starting to use injection drugs or date of first positive serologic test. Hypertension was defined by clinical diagnosis or two consecutive visits with SBP at least 140 mmHg or DBP at least 90 mmHg. Diabetes was defined by clinical diagnosis or two consecutive visits with either fasting serum glucose at least 7 mmol/l or random serum glucose at least 11.1 mmol/l. Liver fibrosis was measured using the aspartate aminotransferase-to-platelet ratio index (APRI) with values at least 1.5 indicative of significant fibrosis (≥ F2) [29 (link)].

Rossi C., Saeed S., Cox J., Vachon M.L., Martel-laferrière V., Walmsley S.L., Cooper C., Gill M.J., Hull M., Moodie E.E, & Klein M.B. (2018). Hepatitis C virus cure does not impact kidney function decline in HIV co-infected patients. AIDS (London, England), 32(6), 751-759.

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Real-world Hepatitis C Treatment in Taiwan

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The National Health Insurance Administration of Taiwan began to reimburse DAAs in January 2017, and there are currently no limitations for treating CHC patients with DAAs. The treatment regimens and strategies conformed to the regional consensus [13 (link),14 (link)] and regulations of the Health and Welfare Department of Taiwan [15 ]. The treatment regimens in the current study included sofosbuvir/ledipasvir, sofosbuvir/velpatasvir, elbasvir/grazoprevir, and glecaprevir/pibrentasvir. HCV antibodies were detected by a third-generation enzyme immunoassay (Abbott Laboratories, North Chicago, IL, USA). HCV RNA and the genotypes were determined using a real-time PCR assay (RealTime HCV; Abbott Molecular, Des Plaines IL, USA; detection limit: 12 IU/mL) [16 (link)]. The institutional review board of Kaohsiung Medical University Hospital approved the protocols, which followed the guidelines of the International Conference on Harmonization for Good Clinical Practice (IRB approval numbers: KMUHIRB-F(I)-20170053, KMUHIRBE(I)-20200245).

Huang C.F., Wu P.F., Yeh M.L., Huang C.I., Liang P.C., Hsu C.T., Hsu P.Y., Liu H.Y., Huang Y.C., Lin Z.Y., Chen S.C., Huang J.F., Dai C.Y., Chuang W.L, & Yu M.L. (2020). Scaling up the in-hospital hepatitis C virus care cascade in Taiwan. Clinical and Molecular Hepatology, 27(1), 136-143.

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HCV Genomic Analysis Protocol

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Genomic DNAs were extracted from 100 µl of sera samples using genomic DNA isolation kit (Qiagen, England) and DNAs samples were dissolved in 70 µl distilled deionized water and kept at -20ºC till polymerase chain reactions (PCR) and restriction fragment length polymorphism (RFLP) tests. HCV RNA was extracted from 100 µl of serum using a viral RNA TRIZOLE Kit (Bioline, England), according to manufacture instructions. HCV RNA samples were dissolved in 70 µl elusion buffer. According to (Vermehren et al., 2011), HCV RNA was quantified by a real-time polymerase chain reaction assay (detection limit: 50 IU/ml; Real-time HCV; Abbott Molecular, Des Plaines IL, USA). Assay protocols, cut-offs and result interpretations were carried out according to the manufacturer’s instructions. The genotype of the amplified cDNA was determined by reverse hybridization using the line probe assay (INNO LiPA HCV II kit, Innogenetics Q2, Ghent, Belgium).

Koutb F., Abdel-Rahman S., Hassona E, & Haggag A. (2017). Association of C-myc and p53 Gene Expression and Polymorphisms with Hepatitis C (HCV) Chronic Infection, Cirrhosis and Hepatocellular Carcinoma (HCC) Stages in Egypt. Asian Pacific Journal of Cancer Prevention : APJCP, 18(8), 2049-2057.

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Hepatitis C Antibody and RNA Detection

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The anti-HCV antibody was detected using a third-generation, commercially available enzyme-linked immunosorbent assay kit (AxSYM 3.0; Abbott Laboratories, Chicago, IL). The presence of serum HCV RNA was evaluated using a standardized automated quantitative reverse-transcription polymerase chain reaction (COBAS AMPLICOR HCV Test, version 2.0; Roche, Branchburg, NJ; detection limit: 50 IU/mL) before 2011 or a real-time polymerase chain reaction assay (RealTime HCV; Abbott Molecular, Des Plaines IL; detection limit: 12 IU/mL) after 2011.^{39 (link)}

Yu M.L., Yeh M.L., Tsai P.C., Huang C.I., Huang J.F., Huang C.F., Hsieh M.H., Liang P.C., Lin Y.H., Hsieh M.Y., Lin W.Y., Hou N.J., Lin Z.Y., Chen S.C., Dai C.Y., Chuang W.L, & Chang W.Y. (2015). Huge Gap Between Clinical Efficacy and Community Effectiveness in the Treatment of Chronic Hepatitis C: A Nationwide Survey in Taiwan. Medicine, 94(13), e690.

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HCV RNA and Genotype Testing Protocol

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The serum HCV RNA at baseline, treatment W4, W8, W12, and W24 were determined by standardized automated real-time PCR assays (Roche Cobas Taqman HCV v2.0 Roche Diagnostics GmbH, Mannheim, Germany) with a detection limit of 15 IU/mL; or RealTime HCV (Abbott Molecular, Des Plaines IL, USA) with a detection limit of 12 IU/ml).[15 (link)]
HCV genotype was determined by a reverse hybridization assay (Versant HCV Genotype 2.0 assay, Siemens Healthcare Diagnostics, Illinois) or a real-time PCR assay (Abbott RealTime HCV Genotype II; Abbott Molecular, Des Plaines IL, USA).[16 (link)] The sustained virological response (SVR) was defined as seronegativity of HCV RNA throughout 24 weeks of post-treatment follow-up period.

Yang C.C., Tsai W.L., Su W.W., Huang C.F., Cheng P.N., Lo C.C., Tseng K.C., Mo L.R., Wang C.H., Hsu S.J., Lai H.C., Su C.W., Liu C.J., Peng C.Y, & Yu M.L. (2015). Rapid Prediction of Treatment Futility of Boceprevir with Peginterferon-Ribavirin for Taiwanese Treatment Experienced Hepatitis C Virus Genotype 1-Infected Patients. PLoS ONE, 10(9), e0137852.

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HCV Genotyping and Treatment Response

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We used a qualitative real-time polymerase chain reaction (Cobas Amplicor Hepatitis C Virus Test 2.0, Roche, Branchburg, NJ)^{[27 (link)]} and a quantification branched DNA assay (Versant HCV RNA 3.0, Bayer, Tarrytown, NJ) with a detection limit of 615 IU/mL or real-time HCV (Abbott Molecular, Des Plaines, IL) with a detection limit of 12 IU/mL^{[28 (link)]} to detect serum HCV RNA. The HCV genotypes were determined using the Okamoto method^{[29 (link)]} or a real-time PCR assay (Abbott real-time HCV Genotype II; Abbott Molecular). Patients with mixed genotypes infection including G1 were classified as G1; those mixed with G2 but without G1 were classified as G2; the others were not included in the current study due to limited case numbers. The liver histology obtained within 1 year before antiviral therapy was classified and staged according to the scoring system described by Scheuer.^{[30 (link)]} A negativity of HCV RNA on a 24-week after treatment follow-up period was defined as achieved successful therapy, SVR. The IL28B rs8099917 genotype was determined by the method described in the previous study.^{[31 (link)]}

Tsai P.C., Liu T.W., Tsai Y.S., Ko Y.M., Chen K.Y., Lin C.C., Huang C.I., Liang P.C., Lin Y.H., Hsieh M.Y., Hou N.J., Huang C.F., Yeh M.L., Lin Z.Y., Chen S.C., Dai C.Y., Chuang W.L., Huang J.F, & Yu M.L. (2017). Identification of groups with poor cost-effectiveness of peginterferon plus ribavirin for naïve hepatitis C patients with a real-world cohort and database. Medicine, 96(22), e6984.

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Diagnosis and Monitoring of HCV Infection

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Anti-HCV antibody was determined by the third generation, commercially available immunoassay (Ax SYM HCV III; Abbott Laboratories, North Chicago, IL). HCV RNA viral loads and genotype were determined by real-time PCR assays [RealTime HCV; Abbott Molecular, Des Plaines IL, United States; detection limit: 12 IU/mL])[22 (link)]. Liver cirrhosis was defined by the presence of clinical, radiological, endoscopic or laboratory evidence of cirrhosis and/or portal hypertension or fibrosis-4 index (FIB-4) (> 6.5). Laboratory data monitoring and assessment of side effects were performed at treatment wk 2, 4, 8 and end-of-treatment (EOT), and 12 wk after EOT.
The primary endpoint was sustained virological response (SVR12, defined as undetectable HCV RNA throughout 12 wk of the post-treatment follow-up period).

Chen C.T., Lu M.Y., Hsieh M.H., Tsai P.C., Hsieh T.Y., Yeh M.L., Huang C.I., Tsai Y.S., Ko Y.M., Lin C.C., Chen K.Y., Wei Y.J., Hsu P.Y., Hsu C.T., Jang T.Y., Liu T.W., Liang P.C., Hsieh M.Y., Lin Z.Y., Huang C.F., Huang J.F., Dai C.Y., Chuang W.L., Shih Y.L, & Yu M.L. (2022). Outreach onsite treatment with a simplified pangenotypic direct-acting anti-viral regimen for hepatitis C virus micro-elimination in a prison. World Journal of Gastroenterology, 28(2), 263-274.

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