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Chapter 36 : Molecular Detection and Characterization of Hepatitis C Virus

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Abstract:

Hepatitis C virus (HCV) and molecular methods are inextricably linked, in history and in clinical practice. This virus was the first to be identified with molecular methods, and nucleic acid tests (NATs) have become fundamentally important in the diagnosis and therapeutic management of the chronic infections it causes. Rates of spontaneous virus clearance are highest in acutely infected individuals who manifest signs of hepatitis, suggesting that an early immune response may be protective. Risk factors for disease progression include diseases or behaviors that induce additional hepatic injury (such as concomitant hepatitis B virus [HBV] infection and alcohol consumption) or impair antiviral immunity (such as HIV infection). Treatment for HCV infection has undergone significant evolution in regard to available agents, duration, and dosing strategy. Important concepts in regard to testing for acute HCV infection are that (i) serology and RNA assessment should both be performed and (ii) testing should be undertaken at multiple time points before exclusion of the diagnosis. Given the asymptomatic nature of most acute HCV infections, the majority of patients will present symptomatically, in the chronic phase. The approval of STAT-C drugs has the potential to vastly improve treatment efficacy and concomitantly impose a new paradigm for therapeutic monitoring. The effect of telaprevir on the performance of viral load testing and the necessity for additional tests such as resistance assays remains to be determined.

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36

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TaqMan Real-Time PCR Assay
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Restriction Fragment Length Polymorphism
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Reverse Transcriptase PCR
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Image of FIGURE 1
FIGURE 1

HCV genome and protein coding scheme. nuc, nucleotide; pos, position; aa, amino acid; gp, glycoprotein. Numbering according to references and .

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 2
FIGURE 2

Clinical features of acute hepatitis C following exposure to low viral inoculum such as occupational needlestick exposure or community-based exposure ( ). Characteristics following higher-dose exposure (transfusion with contaminated blood products) may be different. Intermittent viremia phase estimated from needlestick exposure ( ). Kinetics of other characteristics derived from seroconversion panels ( ). ALT, alanine aminotransferase. Dashed lines indicate potential viremia patterns as defined by HCV RNA levels in peripheral blood. Adapted from reference .

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 3
FIGURE 3

Liver disease progression in chronic hepatitis C. Factors listed in notched arrow accelerate histologic progression of disease. HCC, hepatocellular carcinoma.

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 4
FIGURE 4

Evolution of treatment and therapeutic efficacy (% SVR) for chronic hepatitis C. Years represent date of first report (IFN alfa, triple therapy) or randomized clinical trial report (IFN alfa/RBV, pegIFN alfa/RBV). Triple therapy, pegIFN/RBV/telaprevir (protease inhibitor) for 28 days, then treatment for up to 44 additional weeks with pegIFN/RBV. IFN, interferon; RBV, ribavirin.

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 5
FIGURE 5

Therapeutic response definitions.

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 6
FIGURE 6

Individualized chronic hepatitis C therapy algorithms based on genotype, baseline viral load, and response kinetics. Rx wk, treatment week; LVL, low baseline viral load (<400,000 IU/ml); HVL, high baseline viral load (≥400,000 IU/ml); UD, undetectable; +, detectable HCV RNA. Adapted from reference .

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 7
FIGURE 7

Relation between HCV RNA clearance pattern and EOT response, relapse rate, and SVR. UD, undetectable HCV RNA. Data from reference .

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 8
FIGURE 8

Evolution of proposed diagnostic testing algorithms for acute hepatitis C following known exposure. Alanine aminotransferase (ALT) measurement recommended to document biochemical evidence of hepatitis. Enzyme immunoassay (EIA) for detection of anti-HCV antibodies. NAT, qualitative or highly sensitive quantitative (real-time PCR-based) nucleic acid test. Derived from references , and .

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Image of FIGURE 9
FIGURE 9

Monitoring response to chronic hepatitis C treatment: timing and appropriate tests. Gray boxes, test with quantitative assay; black boxes, test with qualitative assay or sensitive (realtime PCR-based) quantitative assay; white boxes, no testing indicated; BL, baseline.

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
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Tables

Generic image for table
TABLE 1

HCV RNA qualitative tests

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
Generic image for table
TABLE 2

HCV RNA quantitative tests

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36
Generic image for table
TABLE 3

HCV RNA genotyping tests

Citation: Forman M, Valsamakis A. 2011. Molecular Detection and Characterization of Hepatitis C Virus, p 557-577. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch36

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