Evaluation Study of RDTs Detecting Antibodies Against HCV

Evaluation Study of Rapid Diagnostic Tests (RDTs) Detecting Antibodies Against Hepatitis C Virus

The study aims to evaluate 13 different HCV RDTs (10 on-market, 3 under development) for their diagnostic performance and operational characteristics in archived EDTA plasma samples, originating from patients from different geographical regions (Nigeria, Georgia, Cambodia, Belgium) and with or without HIV co-infection.

Study Overview

• Status: Completed
• Conditions: Hepatitis C
• Intervention / Treatment: Diagnostic test: 13 Rapid Diagnostic Tests and reference tests

Detailed Description

Background and rationale:

Screening of past exposure to Hepatitis C Virus (HCV) infection is done by detection of HCV specific antibodies. In Low and Middle Income Countries (LMICs), where equipped laboratories and trained staff are limited, Rapid Diagnostic Tests (RDTs) are widely used for HCV screening. Although many RDTs are available on the market, only some of them received CE-IVD marking and only two have been validated by WHO Pre-Qualification (PQ). More quality-assured tests are needed to establish effective screening programmes in LMICs.

Furthermore, an important research gap is the lack of studies on the impact of HIV positivity on RDT performance, as it is estimated that 2-15% of people living with HIV are co-infected with HCV.

The evaluation of RDT performance on clinical samples collected in different geographic regions as well as from HIV co-infected individuals, would allow to identify tests with a performance meeting or having the potential to meet WHO quality standards.

Concept:

This is a multicenter laboratory evaluation study using archived, frozen plasma samples.

Sensitivity and specificity of RDTs will be measured against a composite reference standard that consists of two WHO prequalified Enzyme Immunoassays (EIAs) (Murex Anti-HCV EIA version 4.0, Fujirebio Innotest HCV Ab IV) and a Line Immunoassay (LIA) (MP Diagnostics HCV blot 3.0). Samples are assigned as anti-HCV negative or anti-HCV positive based on the results of all three assays.

RDT results will be read by three independent readers to evaluate inter-reader variability (differences in visual interpretation, i.e. presence or absence of test and control line).

For each RDT, two independently produced lots will be tested for each sample to assess lot-to-lot variability (differences in RDT result for the same sample). Furthermore, rate of invalid runs will be assessed and a technical appraisal is completed for each RDT.

Primary objective:

1.1 Evaluation of sensitivity and specificity of anti-HCV RDTs in archived plasma samples, collected from HCV-infected and HCV-uninfected individuals not co-infected with HIV, measured against the composite reference standard composed of two Enzyme Immunoassays (EIAs) and a Line Immunoassay (LIA).

1.2 Evaluation of sensitivity and specificity of anti-HCV RDTs in archived plasma samples, collected from HCV-infected and HCV-uninfected individuals who are all co-infected with HIV, measured against the composite reference standard composed of two EIAs and a LIA.

Secondary objectives:

2.1 Evaluation of sensitivity and specificity of anti-HCV RDTs in archived plasma samples, collected from HCV-infected and HCV-uninfected individuals, both co-infected and not with HIV, measured against the composite reference standard composed of two EIAs and a LIA.

2.2 Evaluation of operational characteristics of anti-HCV RDTs: inter-reader variability; lot-to-lot variability; rate of invalid runs 2.3 Technical appraisal of each RDT product per manufacturer

• Study Type: Observational
• Enrollment (Actual): 1710

Contacts and Locations

Study Locations

• Belgium
  • Antwerp, Belgium, 2000
    • Institute of Tropical Medicine
• Georgia
  • Tbilisi, Georgia, 0198
    • National Center for Disease Control & Public Health/Lugar Center
• Nigeria
  • Lagos, Nigeria
    • Nigerian Institute of Medical Research

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 16 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Probability Sample

Study Population

Samples tested in Belgium originate from Cambodia (80%) and Belgium (20%). Cambodian samples are leftovers from samples collected during a cross-sectional study of hepatitis C among people living with HIV (De Weggheleire, A. 2017); samples in Belgium were collected and archived during routine patient care visits to the HIV clinic.

Samples in Nigeria originate from routine diagnostic activities at NIMR as well as the AIDS Prevention Initiative Nigeria (APIN) PEPFAR Program Sample/Data Bank setup at NIMR and the Lagos University Teaching Hospital.

Samples tested in Georgia are leftover samples from a previous FIND HCV study (Reipold, E.I., 2019), as well as archived routine diagnostic samples from the site.

Percent sample distribution per site:

Belgium: 38% Nigeria: 41% Georgia: 21%

Description

Inclusion Criteria of archived samples:

• Non-hemolytic plasma samples with EDTA used as anticoagulant
• Sample were frozen at -20°C or lower on the day of processing and stored at -20°C or lower until they are used in this study
• Samples pre-characterized for, HCV, HIV serology status using assays routinely used at the sites and approved for diagnostic use by a local health authority. If available, samples should also be characterized for HBV status.
• Samples taken from subjects aged ≥18 years
• Availability of informed consent to use the sample in future research

Exclusion Criteria:

- Samples not stored correctly

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Retrospective

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
HCV-only infected; Archived frozen plasma samples from individuals that were characterised to be HCV-antibody positive or HCV-antibody negative ("HCV-only infected"). These samples are characterised for their HIV status (negative).	Diagnostic test: 13 Rapid Diagnostic Tests and reference tests; Rapid diagnostic tests: HCV antibody test (under development); bioLytical Laboratories; DPP® HCV (under development); Chembio Diagnostic Systems; HCV-Ab Rapid Test; Beijing Wantai Biological Pharmacy Enterprise; Rapid Anti-HCV Test; InTec; First Response HCV Card Test; Premire Medical Corporation; Signal HCV Ver 3.0; Arkray healthcare; TRI DOT HCV; J. Mitra & Co; Triplex HIV, HCV, HBsAg; Biosynex SA; Standard Q HCV Ab; SD Biosensor; HCV Hepatitis Virus Antibody Test; Artron Laboratories; SD Bioline HCV; Abbott Diagnostics; OraQuick HCV; OraSure; Care Start HCV Rapid Test (under development); Access Bio Reference tests: Enzyme Immunoassays (EIAs): Murex Anti-HCV EIA version 4.0; Fujirebio Innotest HCV Ab IV Line Immunoassay (LIA): MP Diagnostics HCV blot 3.0
HCV/HIV co-infected; Archived frozen plasma samples from HCV-positive or HCV-negative individuals who are HIV infected ("HCV/HIV co-infected").	Diagnostic test: 13 Rapid Diagnostic Tests and reference tests; Rapid diagnostic tests: HCV antibody test (under development); bioLytical Laboratories; DPP® HCV (under development); Chembio Diagnostic Systems; HCV-Ab Rapid Test; Beijing Wantai Biological Pharmacy Enterprise; Rapid Anti-HCV Test; InTec; First Response HCV Card Test; Premire Medical Corporation; Signal HCV Ver 3.0; Arkray healthcare; TRI DOT HCV; J. Mitra & Co; Triplex HIV, HCV, HBsAg; Biosynex SA; Standard Q HCV Ab; SD Biosensor; HCV Hepatitis Virus Antibody Test; Artron Laboratories; SD Bioline HCV; Abbott Diagnostics; OraQuick HCV; OraSure; Care Start HCV Rapid Test (under development); Access Bio Reference tests: Enzyme Immunoassays (EIAs): Murex Anti-HCV EIA version 4.0; Fujirebio Innotest HCV Ab IV Line Immunoassay (LIA): MP Diagnostics HCV blot 3.0

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Point estimates of sensitivity and specificity, with 95% confidence intervals, of RDTs for the detection of anti-HCV antibodies in individuals not co-infected with HIV.	Point estimates of sensitivity and specificity, with 95% confidence Intervals based on Wilson's score method, will be computed for all samples HCV+ve/HIV-ve and HCV-ve/HIV-ve; the calculation was performed for all RDT manufacturers and individually for each lot. The estimates were calculated on the overall sample population.	6 months
Point estimates of sensitivity and specificity, with 95% confidence intervals, of RDTs for the detection of anti-HCV antibodies in HIV co-infected individuals.	Point estimates of sensitivity and specificity, with 95% confidence Intervals based on Wilson's score method, were computed for all samples HCV+ve/HIV+ve and HCV-ve/HIV+ve; the calculation was performed for all RDT manufacturers and individually for each lot. The estimates were calculated on the overall sample population.	6 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Point estimates of sensitivity and specificity, with 95% confidence intervals, of RDTs for the detection of anti-HCV antibodies in the overall sample population.	Point estimates of sensitivity and specificity, with 95% confidence intervals based on Wilson's score method, were computed for all samples: HCV+ve/HIV-ve, HCV-ve/HIV-ve, HCV+ve/ HIV+ve and HCV-ve/HIV+ve; the was performed for all RDT manufacturers and individually for each lot. The estimates were calculated on the overall sample population.	6 months
Operational characteristics	Inter-reader variability: Fleiss Kappa Coefficient (κ) of inter-reader variability per RDT; manufacturer lot Lot-to-lot variability: coefficient of lot-to-lot variability (percentage agreement) per RDT manufacturer Rate of invalid runs: Percent of invalid runs per manufacturer lot	6 months
Technical appraisal rating on kit instructions, packaging, labelling and test conduct, on a Likert scale	Averages of likert-scale scores will be calculated and serve to understand the user-friendliness of the RDT; there is no expected outcome, the scale rating is used to collect consistent feedback from users; scale scores are as follows: 1=poor, 2=needs improvement, 3=satisfactory, 4=good, 5=excellent	6 months

Collaborators and Investigators

• Sponsor: Foundation for Innovative New Diagnostics, Switzerland
• Collaborators: Institute of Tropical Medicine, Belgium; Nigerian Institute of Medical Research; Richard Lugar Centre for Public Health Research, Georgia
• Investigators: Study Director: Ranga Sampath, PhD, Foundation for Innovative New Diagnostics, Geneva, Switzerland; Principal Investigator: Rosemary Audu, PhD, Nigerian Institute of Medical Research, Lagos, Nigeria; Principal Investigator: Katrien Fransen, PhD, Institute of Tropical Medicine, Antwerp, Belgium; Principal Investigator: Maia Alkhazashvili, Masters, NCDC Lugar Centre, Tbilisi, Georgia

Publications and helpful links

General Publications

• De Weggheleire A, An S, De Baetselier I, Soeung P, Keath H, So V, Ros S, Teav S, Smekens B, Buyze J, Florence E, van Griensven J, Thai S, Francque S, Lynen L. A cross-sectional study of hepatitis C among people living with HIV in Cambodia: Prevalence, risk factors, and potential for targeted screening. PLoS One. 2017 Aug 23;12(8):e0183530. doi: 10.1371/journal.pone.0183530. eCollection 2017.
• Reipold, E.I., Evaluation of the diagnostic performance of the Xpert® Fingerstick HCV Viral Load (VL) Assay. Manuscript in preparation 2019
• Vetter BN, Reipold EI, Ongarello S, Audu R, Ige FA, Alkhazashvili M, Chitadze N, Vanroye F, De Weggheleire A, An S, Fransen K. Sensitivity and Specificity of Rapid Diagnostic Tests for Hepatitis C Virus With or Without HIV Coinfection: A Multicentre Laboratory Evaluation Study. J Infect Dis. 2022 Aug 26;226(3):420-430. doi: 10.1093/infdis/jiaa389.

Study record dates

Study Major Dates

• Study Start (Actual): September 21, 2018
• Primary Completion (Actual): March 15, 2019
• Study Completion (Actual): March 15, 2019

Study Registration Dates

• First Submitted: March 18, 2019
• First Submitted That Met QC Criteria: July 25, 2019
• First Posted (Actual): July 26, 2019

Study Record Updates

• Last Update Posted (Actual): July 29, 2019
• Last Update Submitted That Met QC Criteria: July 26, 2019
• Last Verified: July 1, 2019

More Information

Terms related to this study

• Keywords: Diagnosis; HIV coinfection; Rapid diagnostic test
• Additional Relevant MeSH Terms: Digestive System Diseases; RNA Virus Infections; Virus Diseases; Infections; Blood-Borne Infections; Communicable Diseases; Liver Diseases; Flaviviridae Infections; Hepatitis, Viral, Human; Hepatitis; Hepatitis C
• Other Study ID Numbers: 8162-2/1

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No