Tools for the diagnosis of hepatitis C virus infection and hepatic fibrosis staging

Table 1 Main commercial immunoassays to detect anti-hepatitis C virus antibodies approved for in vitro diagnostics

Analyzer and manufacturer	Assay principle	Solid phase	HCV antigens	Reaction sample volume (μL)	Time of reaction (min)	IVD registration
Architect i2000SR, Abbott Laboratories	CMIA	Paramagnetic particles	HCr43 (Core and NS3), c100-3 (NS4A)	20	20	FDA, CE
AxSYM, Abbott Laboratories	MEIA	Paramagnetic particles	HCr43 (Core), c200 (NS3), c100-3 (NS4A)	33	30	FDA, CE
LiaisonXL, DiaSorin	CLIA	Paramagnetic particles	Core, NS3, NS4	25	46	CE
VITROS ECi, VITROS 3600, Ortho-Clinical Diagnostics	CLIA	Microwell	c22-3 (Core), c200 (NS3 and NS4), NS5	20	55	FDA, CE
Elecsys, Roche Diagnostics	ECLIA	Paramagnetic particles	Core, NS3, NS4	40	18	FDA, CE
ADVIA Centaur, Siemens	CLIA	Magnetic particles	c22-3 (Core), NS3, c200, NS5	10	58	FDA, CE

IVD: Certified as in vitro diagnostic test or device; CE: Conformité Européenne (European Union); FDA: Food and Drug Administration (United States of America); ECLIA: Electrochemiluminescence immunoassay; CMIA: Chemiluminescent microparticle immunoassay; CLIA: Chemiluminescence immunoassay; HCV: Hepatitis C viru; MEIA: Microparticle capture enzyme immunoassay.

Table 2 Hepatitis C virus-RNA qualitative assays approved for in vitro diagnostics

Assay and manufacturer	Method	Reaction sample volume (μL)	Lower limit of detection (IU/mL)	Instrumentation for automated processing	IVD registration
COBAS® AMPLICOR HCV Test v2.0, Roche Molecular Systems	RT-PCR	500	50 (plasma) 60 (serum)	COBAS® AMPLICOR® Analyzer (amplification and detection)	CE, FDA, Japan, Canada
COBAS® AmpliPrep/COBAS® AMPLICOR HCV Test v2.0, Roche Molecular Systems	RT-PCR	250	50 (plasma) 60 (serum)	COBAS® AmpliPrep (extraction), COBAS® AMPLICOR® Analyzer (amplification and detection)	CE, FDA, Canada
COBAS® AmpliPrep/COBAS® TaqMan® HCV Qualitative Test v2.0, Roche Molecular Systems	Real-time RT-PCR	650	15	Fully automated: cobas p 630 Instrument (primary tube handling), COBAS® AmpliPrep (extraction and MM setup), COBAS® TaqMan® Analyzer or the COBAS® TaqMan® 48 Analyzer (amplification and detection)	CE, FDA
APTIMA HCV RNA Qualitative Assay1, Hologic - Gen-Probe	TMA	500	5.3	Not automated. PANTHER System’s functionality currently in development	FDA
VERSANT® HCV RNA Qualitative Assay, Siemens	TMA	50	5.3	TMA modules (TCS, luminometer HC+, etc.)	CE, FDA

¹The performance of this assay has not been demonstrated for monitoring hepatitis C virus (HCV) infected patients. IVD: Certified as in vitro diagnostic test or device; CE: Conformité Européenne (European Union); FDA: Food and Drug Administration (United States of America); RT-PCR: Reverse transcription-polymerase chain reaction; MM: Master mix.

Table 3 Hepatitis C virus-RNA quantitative assays approved for in vitro diagnostics

Assay and manufacturer	Method	Reaction sample volume (μL)	Lower limit of detection (IU/mL)	Linear range of quantification (IU/mL)	Instrumentation for automated processing	IVD registration
Abbott RealTime HCV, Abbott Molecular	Real-time RT-PCR	500	12	12 - (1 × 108)	m2000sp (extraction and assay setup), m2000rt (amplification and detection)	CE, FDA
artus HCV RG RT-PCR Kit, Qiagen	Real-time RT-PCR	500	34	65 - (1 × 106)	Manual extraction (QIAamp DSP Virus Kit) and assay setup, Rotor-Gene Q (amplification and detection)	CE
artus HCV QS-RGQ Kit, Qiagen	Real-time RT-PCR	1000	21	35 - (1.77 × 107)	QIAsymphony RGQ: QIAsymphony SP (extraction), QIAsymphony AS (assay setup), Rotor-Gene Q (amplification and detection)	CE
COBAS® AmpliPrep/COBAS® TaqMan® HCV Test, Roche Molecular Systems	Real-time RT-PCR	850	15	43 - (6.9 × 107)	COBAS® AmpliPrep (extraction), COBAS® TaqMan® Analyzer or COBAS® TaqMan® 48 Analyzer (amplification and detection)	CE, FDA, Canada, Japan
COBAS® AmpliPrep/COBAS® TaqMan® HCV Quantitative Test v2.0, Roche Molecular Systems	Real-time RT-PCR	650	15	15 - (1 × 108)	Fully automated: cobas p 630 (primary tube handling), COBAS® AmpliPrep (extraction), COBAS® TaqMan® Analyzer or COBAS® TaqMan® 48 Analyzer (amplification and detection)	CE, FDA
COBAS® TaqMan® HCV Test v2.0 for use with the High Pure System, Roche Molecular Systems	Real-time RT-PCR	500	10	25 - (3.9 × 108) (CE)	Manual extraction (High Pure System Viral Nucleic Acid Kit), COBAS® TaqMan® Analyzer (amplification and detection)	CE, FDA
				25 - (3.0 × 108) (FDA)
VERSANT® HCV RNA 1.0 Assay (kPCR), Siemens	Real-time RT-PCR	500	15	15 - (1.0 × 108)	VERSANT® kPCR Molecular System Sample	CE
					Preparation (SP) Module and Amplification/Detection (AD) Module
VERSANT® HCV RNA 3.0 Assay, Siemens	bDNA	50	6151	615 - (7.7 × 106)	No nucleic acid extraction needed. System 340 bDNA Analyzer (S340) (US) or VERSANT™ 440 Molecular System (CE)	CE, FDA

¹The intended use of this assay is limited to the measurement of hepatitis C virus (HCV) viral loads at baseline and after 12 wk of therapy. IVD: Certified as in vitro diagnostic test or device; CE: Conformité Européenne (European Union); FDA: Food and Drug Administration (United States of America); kPCR: Real-time kinetic polymerase chain reaction; bDNA: Branched DNA; RT-PCR: Reverse transcription-polymerase chain reaction.

Table 4 Hepatitis C virus genotyping assays approved for in vitro diagnostics

Assay and manufacturer	Method	Reaction sample volume (μL)	Lower limit of detection (IU/mL)	Target	Instrumentation	IVD registration
Abbott HCV Genotype II, Abbott Molecular	Real-time RT-PCR	500	500	5'UTR	m2000sp (extraction and assay setup), m2000rt (amplification and detection)	CE, FDA
				NS5B
LINEAR ARRAY Hepatitis C Virus Genotyping Test, Roche Molecular Systems	RT-PCR and hybridization	200	500	5'UTR	AMPLICOR® and COBAS® AMPLICOR HCV Test v2.0 (amplification and detection). Manual hybridization.	CE
VERSANT HCV Genotype 2.0 Assay Line Probe Assay (LiPA), Siemens	RT-PCR and hybridization	500–1000	2000	5'UTR core	Conventional PCR instrument, Auto-LiPA 48 or AutoBlot 3000H automated Systems (hybridization), LiPA Scan Software CD (reading)	CE

IVD: Certified as in vitro diagnostic test or device; CE: Conformité Européenne (European Union); FDA: Food and Drug Administration (United States of America); RT-PCR: Reverse transcription-polymerase chain reaction.

Table 5 Serum-based tests recommended in the clinical practice for the detection of significant fibrosis

Serum-based tests	Parameters considered	Ref.
Tests combining direct markers of liver fibrosis
Enhanced liver Fibrosis test (ELF™)	HA, PIIINP, TIMP-1, age	[88]
MP3™	MMP-1, PIIINP	[89]
Fibrospect II™	HA, TIMP-1, alpha-2-macroglobulin	[90]
Tests including indirect markers of liver fibrosis
AST to platelet ratio index (APRI)	AST, platelet count	[91]
AST/ALT ratio	AST, ALT	[92]
Forns index	GGT, platelet count, cholesterol, age	[93]
Tests including combinations of direct and indirect markers of fibrosis
Hepascore™	Bilirubin, GGT, HA, alpha-2-macroglobulin, gender, age	[94]
Fibrometer™	HA, AST, platelet count, prothrombin index, alpha-2-macroglobulin, urea, age	[95]
FibroTest™	alpha-2-macroglobulin, haptoglobin, apolipoprotein A1, GGT, total bilirubin	[96]

HA: Hyaluronic acid; PIIINP: Pro-collagen III amino-terminal peptide; TIMP-1: Tissue inhibitor of matrix metalloproteinase-1; MMP-1: Matrix metalloproteinase-1; AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; GGT: Gamma-glutamyltransferase.