HCV-RNA Decline Research Articles

546 Early HCV-RNA decline in peripheral blood mononuclear cells predicts SVR versus relapse in chronic hepatitis C patients treated with pegylated interferon-α plus ribavirin

546 Early HCV-RNA decline in peripheral blood mononuclear cells predicts SVR versus relapse in chronic hepatitis C patients treated with pegylated interferon-α plus ribavirin

Perioperative viral kinetics: New insights or the Emperor's new clothes?

Perioperative viral kinetics: New insights or the Emperor's new clothes?

Pharmacokinetics, pharmacodynamics, and hepatitis C viral kinetics during antiviral therapy: The null responder

Our aim was to study the effect of ribavirin on viral kinetics and to study the early patterns of response to antiviral therapy of hepatitis C and their correlation with interferon pharmacokinetics and pharmacodynamics. We conducted a randomized, controlled trial of peginterferon alfa-2a with or without ribavirin in interferon naïve patients with HCV genotype 1. HCV RNA levels were measured at frequent intervals during treatment together with serum levels of peginterferon alfa-2a and 2'5'oligoadenylate synthetase (OAS). Of 29 patients treated, 14 had a complete response at day 29 while 15 had a null response (less than 1 log decline in HCV RNA). There were no significant differences between complete and null responders with regard to gender, age, race, body mass index, HCV subtype, baseline HCV RNA levels, serum aminotransferase activities, stage of fibrosis, peak OAS, or interferon levels. Mean serum IFN levels at day 29 and OAS levels at day 3 were no different between complete and null responders (15,525 vs. 30,768 pg/ml and 2,044 vs. 2,323 pM/hr, respectively, P = n.s.). Addition of ribavirin to pegylated IFN did not significantly affect day 29 IFN levels or Day 3 OAS levels. In conclusion, null responders to either peginterferon alone or in combination with ribavirin have little or no decrease in serum HCV RNA early in therapy and rarely go on to achieve sustained virologic response and have no apparent host differences than complete responders, suggesting that interferon resistance in these patients may be virally rather than host-determined.

O.064 Antiviral treatment for hepatitis C virus infection in the real practice: A nationwide Italian experience (the PROBE study)

The combination of pegylated interferons (PEG-IFNs) and ribavirin represents the standard of care for the treatment of chronic HCV-infected patients, yet with a success rate around 50% in genotypes 1 and 4, high costs and side effects. Therefore, early prediction of sustained virological response (SVR) is a relevant issue for HCV-patients. We evaluated the association between SVR and decline of HCV-RNA at 48 h in a prospective cohort of 145 HCV-patients treated with PEG-IFNs and ribavirin (males = 69.1%; genotypes 1/4 = 51.0%; HIV-1 coinfected = 6.7%). SVR was obtained in 65.5% of patients, while 16.6% experienced relapse and 17.9% no response. The first-phase of HCV-RNA decline clearly differentiated patients with SVR from relapsers and non-responders, independently of genotype (P < 0.001). In univariate and multivariate analyses, different infralogaritmic thresholds of HCV-RNA decay at 48 h were tested, observing the highest predictive potential at 0.5 log: decays above this threshold showed a 76.2% negative predictive value for SVR, whereas decays >0.5 log indicated a 6.8 odds ratio (95% C.I.: 2.0–23.2) for SVR after controlling for genotype, baseline viremia, adherence to therapy and HIV coinfection. Decays beyond the 0.5 log threshold were also strongly associated with and highly predictive of early virological response (95.0% positive predictive value, P < 0.001).

P.250 Australian trial in acute hepatitis C: preliminary findings

In patients with chronic hepatitis C virus (HCV) infection scheduled for a 48-week treatment period, premature discontinuation of treatment was previously recommended if HCV-RNA levels remained detectable at week 24 of therapy. Considering the number of side effects and treatment costs, measurement of initial viral decline during therapy may identify virologic nonresponse earlier than 24 weeks. We retrospectively analyzed 260 European patients treated with standard or pegylated interferon alfa (IFN-α) and ribavirin for 24 to 48 weeks. Early prediction of virologic response by HCV-RNA decline at weeks 4 and 12 (Versant Quantitative [branched DNA (bDNA) 3.0]; Bayer Diagnostics, Emeryville, CA; and Qualitative [transcription-mediated amplification (TMA)] HCV RNA assay; Bayer Diagnostics) as well as clinical, biochemical, virologic, and histologic baseline parameters were analyzed by logistic regression and receiver operating characteristic (ROC) curves. A viral load at treatment week 4 above 450,000 IU/mL and at week 12 above 30,000 IU/mL was 100% predictive for virologic nonresponse in all patients. From multivariate logistic regression analysis of all patients, independent predictors for sustained virologic response were: genotypes 2 and 3 (P < .0001), a low baseline gamma-glutamyltransferase (GGT) level (P < .0001), a high baseline alanine aminotransferase level (P = .002), and a low baseline viral load (P = .04). None of the latter 3 factors were predictive for sustained virologic response when analysis was restricted to the subgroup of genotypes 2– and 3–infected patients. In conclusion, virologic nonresponse can be predicted early at week 12 of treatment independent from the applied therapeutic regimen based on a cutoff level for HCV RNA of 30,000 IU/mL. This algorithm recognizes 53.7% of nonresponders previously identified at week 24 of treatment. (HEPATOLOGY 2003;37:600-609.)

P.251 Long term outcome of chronic hepatitis C patients with sustained virological response at 6 months after the end of treatment

In patients with chronic hepatitis C virus (HCV) infection scheduled for a 48-week treatment period, premature discontinuation of treatment was previously recommended if HCV-RNA levels remained detectable at week 24 of therapy. Considering the number of side effects and treatment costs, measurement of initial viral decline during therapy may identify virologic nonresponse earlier than 24 weeks. We retrospectively analyzed 260 European patients treated with standard or pegylated interferon alfa (IFN-α) and ribavirin for 24 to 48 weeks. Early prediction of virologic response by HCV-RNA decline at weeks 4 and 12 (Versant Quantitative [branched DNA (bDNA) 3.0]; Bayer Diagnostics, Emeryville, CA; and Qualitative [transcription-mediated amplification (TMA)] HCV RNA assay; Bayer Diagnostics) as well as clinical, biochemical, virologic, and histologic baseline parameters were analyzed by logistic regression and receiver operating characteristic (ROC) curves. A viral load at treatment week 4 above 450,000 IU/mL and at week 12 above 30,000 IU/mL was 100% predictive for virologic nonresponse in all patients. From multivariate logistic regression analysis of all patients, independent predictors for sustained virologic response were: genotypes 2 and 3 (P < .0001), a low baseline gamma-glutamyltransferase (GGT) level (P < .0001), a high baseline alanine aminotransferase level (P = .002), and a low baseline viral load (P = .04). None of the latter 3 factors were predictive for sustained virologic response when analysis was restricted to the subgroup of genotypes 2– and 3–infected patients. In conclusion, virologic nonresponse can be predicted early at week 12 of treatment independent from the applied therapeutic regimen based on a cutoff level for HCV RNA of 30,000 IU/mL. This algorithm recognizes 53.7% of nonresponders previously identified at week 24 of treatment. (HEPATOLOGY 2003;37:600-609.)

P.249 Recurrence or persistence of HCV RNA detectable only by TMA during antiviral therapy

In patients with chronic hepatitis C virus (HCV) infection scheduled for a 48-week treatment period, premature discontinuation of treatment was previously recommended if HCV-RNA levels remained detectable at week 24 of therapy. Considering the number of side effects and treatment costs, measurement of initial viral decline during therapy may identify virologic nonresponse earlier than 24 weeks. We retrospectively analyzed 260 European patients treated with standard or pegylated interferon alfa (IFN-α) and ribavirin for 24 to 48 weeks. Early prediction of virologic response by HCV-RNA decline at weeks 4 and 12 (Versant Quantitative [branched DNA (bDNA) 3.0]; Bayer Diagnostics, Emeryville, CA; and Qualitative [transcription-mediated amplification (TMA)] HCV RNA assay; Bayer Diagnostics) as well as clinical, biochemical, virologic, and histologic baseline parameters were analyzed by logistic regression and receiver operating characteristic (ROC) curves. A viral load at treatment week 4 above 450,000 IU/mL and at week 12 above 30,000 IU/mL was 100% predictive for virologic nonresponse in all patients. From multivariate logistic regression analysis of all patients, independent predictors for sustained virologic response were: genotypes 2 and 3 (P < .0001), a low baseline gamma-glutamyltransferase (GGT) level (P < .0001), a high baseline alanine aminotransferase level (P = .002), and a low baseline viral load (P = .04). None of the latter 3 factors were predictive for sustained virologic response when analysis was restricted to the subgroup of genotypes 2– and 3–infected patients. In conclusion, virologic nonresponse can be predicted early at week 12 of treatment independent from the applied therapeutic regimen based on a cutoff level for HCV RNA of 30,000 IU/mL. This algorithm recognizes 53.7% of nonresponders previously identified at week 24 of treatment. (HEPATOLOGY 2003;37:600-609.)

P.280 Therapeutic effect of interferon alpha-2b in a case of acute hepatitis C and splenic marginal zone B-cell lymphoma

The aim of the present study was to assess parameters in early phase HCV dynamics for predicting the outcome of interferon (IFN)/ribavirin combination therapy in patients with chronic hepatitis C (CH-C). Sixty-five CH-C patients who received IFN alpha-2b/ribavirin combination therapy were enrolled. The serum levels of HCV RNA 0 h and 3 months after commencing therapy were serially quantified. HCV kinetic parameters such as quantity, ratio of decline, and half-life were analyzed. In genotype 1 patients, both the quantity and the ratio of decline of HCV RNA 24 h after the start of therapy were useful predictors of a poor response. No patients who had serum HCV RNA above 200 KIU/ml 24 h after the start of therapy achieved a sustained viral response (SVR). In genotype 2 patients, conversely, these two parameters were predictors of a sustained viral response. The efficacy of these parameters in predicting the outcome of therapy was comparable to that of the disappearance of HCV RNA from sera at 4 weeks. These results demonstrate that parameters of HCV kinetics 24 h after the start of therapy are useful for the early prediction of outcome in response to IFN alpha-2b/ribavirin combination therapy.

P.239 Hepatitis C antiviral therapy in cirrhotic patients

Amantadine may augment virological response rates to interferon-based therapy in chronic hepatitis C patients. Using a novel design, amantadine was studied in naïve genotype 1 patients treated in combination with peginterferon alfa-2a (40KD)/ribavirin.Patients enrolled in this randomized, placebo-controlled multicenter trial were stratified by single-dose interferon sensitivity (stratum I, 24-h HCV-RNA decline >1.4-log10; II, 0.8–1.39-log10; III, <0.8-log10; a reliable means of identifying nonresponders to interferon/ribavirin) and fibrosis grade (F0/1/2 vs. F3/4) at baseline. All patients received peginterferon alfa-2a (40KD) 180 μg/week plus ribavirin 1000–1200 mg/day and were randomized to receive amantadine 100 mg twice daily (N=114) or placebo (N=95) for 48 weeks.Week-24 virological response rates in strata II and III, the primary outcome, were similar in patients treated with amantadine (63.7%) or placebo (65.7%), as were sustained virological response rates at week 72 (46.5 and 51.6%, respectively). Adverse event profiles were similar and amantadine did not improve health-related quality of life compared with placebo. Interferon sensitivity was the only significant predictor of treatment outcome.Adding amantadine to peginterferon alfa-2a (40KD)/ribavirin combination therapy does not augment virological response rates in genotype 1 patients. Virological response was almost exclusively determined by interferon sensitivity at baseline.

Randomized, double-blind, placebo-controlled study of peginterferon alfa-2a (40KD) plus ribavirin with or without amantadine in treatment-naïve patients with chronic hepatitis C genotype 1 infection

Amantadine may augment virological response rates to interferon-based therapy in chronic hepatitis C patients. Using a novel design, amantadine was studied in naïve genotype 1 patients treated in combination with peginterferon alfa-2a (40KD)/ribavirin. Patients enrolled in this randomized, placebo-controlled multicenter trial were stratified by single-dose interferon sensitivity (stratum I, 24-h HCV-RNA decline >1.4-log10; II, 0.8-1.39-log10; III, <0.8-log10; a reliable means of identifying nonresponders to interferon/ribavirin) and fibrosis grade (F0/1/2 vs. F3/4) at baseline. All patients received peginterferon alfa-2a (40KD) 180 microg/week plus ribavirin 1000-1200 mg/day and were randomized to receive amantadine 100 mg twice daily (N = 114) or placebo (N = 95) for 48 weeks. Week-24 virological response rates in strata II and III, the primary outcome, were similar in patients treated with amantadine (63.7%) or placebo (65.7%), as were sustained virological response rates at week 72 (46.5 and 51.6%, respectively). Adverse event profiles were similar and amantadine did not improve health-related quality of life compared with placebo. Interferon sensitivity was the only significant predictor of treatment outcome. Adding amantadine to peginterferon alfa-2a (40KD)/ribavirin combination therapy does not augment virological response rates in genotype 1 patients. Virological response was almost exclusively determined by interferon sensitivity at baseline.

Developement of Non-Hodgkinʼs Lymphoma during Combination Therapy with Viral Breakthrough

Background: Hepatitis C is the most common chronic blood-borne infection in the United States. Liver is the primary site of infection. HCV can also infect and replicate in peripheral blood mononuclear cells leading to monoclonal proliferation of B-cells. Lymphoproliferative disorders have been associated with HCV including essential mixed cryglobulinemia and non-Hodgkin's lymphoma. 7–20% of patients with NHL have HCV. Antiviral therapy of the HCV infection has been associated with regression of the NHL. To date there are no reports of lymphoma development during antiviral therapy. We are reporting a case of NHL developing during combination therapy with viral breakthrough. Case presentation: A 54 year old Vietnam war veteran with history of IVDA in remission and chronic hepatitis C was referred to the GI-Liver clinic. He was found to have genotype 1b, over 5.000.000 copies per ml viral load and stage 3 fibrosis on liver biopsy. He had history of post traumatic stress disorder, depression and hypertention. He underwent therapy with rebetron for 24 weeks with some biochemical but no virologic response and therapy was terminated. With advanced liver disease he was offered retreatment with pegylated interferon alpha 2 b (1,5 mcg/kg) weekly and ribavirin (10.6 mg/kg) daily. He had 2 log decline in HCV RNA to 21.000 copies per ml at 12 weeks with complete normalization of ALT and complete viral eradication at 24 weeks. At week 47 he presented to his primary care physician with 1-week history of a sore throat and an enlarged, tender lymph node in the right posterior cervical chain. He was given azithromycin with no improvement. A CT of the neck revealed a 2 cm lesion involving the right submandibular gland. FNA of the lesion revealed large cell lymphoma, diffuse, B-type. Repeat viral RNA revealed viral breakthrough with 923,167 copies per ml. The prevalence of breakthrough during interferon therapy ranges between 14% to 21%. The significant viral proliferation during breakthrough in this case has been associated with clonal proliferation of the B-cells and lymphoma development. The pathophysiologic insights, LN biopsy and patient/virus demographics will be discussed.

557 Cost effective prediction of sustained response to IFN α-2B or Peg-IFN α-2B plus ribavirin computing infected cells dynamics by early ALT and HCV-RNA decline

557 Cost effective prediction of sustained response to IFN α-2B or Peg-IFN α-2B plus ribavirin computing infected cells dynamics by early ALT and HCV-RNA decline

High-dose interferon-α2b induction therapy in combination with ribavirin for treatment of chronic hepatitis C in patients with non-response or relapse after interferon-α monotherapy

To evaluate the daily high-dose induction therapy with interferon-alpha2b (IFN-alpha2b) in combination with ribavirin for the treatment of patients who failed with interferon monotherapy and had a relapse, based on the assumption that the viral burden would decline faster, thus increasing the likelihood of higher response rates in this difficult-to-treat patient group. Seventy patients were enrolled in this study. Treatment was started with 10 MU IFN-alpha2b daily for 3 wk, followed by IFN-alpha2b 5 MU/TIW in combination with ribavirin (1 000-1 200 mg/d) for 21 wk. In case of a negative HCV RNA PCR, treatment was continued until wk 48 (IFN-alpha2b 3 MU/TIW+1 000-1 200 mg ribavirin/daily). The dose of IFN-alpha2b or ribavirin was reduced in 16% of patients because of hematologic side effects, and treatment was discontinued in 7% of patients. An early viral response (EVR) was achieved in 60% of patients. Fifty percent of all patients achieved an end-of-treatment response (EOT) and 40% obtained a sustained viral response (SVR). Patients with no response had a significantly lower response rate than those with a former relapse (SVR 30% vs 53%; P = 0.049). Furthermore, lower response rates were observed in patients infected with genotype 1a/b than in patients with non-1-genotype (SVR 28% vs 74%; P = 0.001). As a significant predictive factor for a sustained response, a rapid initial decline of HCV RNA could be identified. No patient achieving a negative HCV-RNA PCR at wk 18 or later eventually eliminated the virus. Daily high-dose induction therapy with interferon-alpha2b is well tolerated and effective for the treatment of non-responders and relapsers, when interferon monotherapy fails. A fast decline of viral load during the first 12 wk is strongly associated with a sustained viral response.

Pegylated Interferon Alfa-2b vs Standard Interferon Alfa-2b, Plus Ribavirin, for Chronic Hepatitis C in HIV-Infected Patients&lt;SUBTITLE&gt;A Randomized Controlled Trial&lt;/SUBTITLE&gt;

Treatment of chronic hepatitis C virus (HCV) infection in human immunodeficiency virus (HIV)-infected patients is a growing concern. Most data on the virologic efficacy and safety of the combination of peginterferon alfa-2b and ribavirin in coinfected patients come from uncontrolled studies. To study the safety and efficacy of peginterferon alfa-2b plus ribavirin vs standard interferon alfa-2b plus ribavirin in HIV-HCV coinfected patients. A multicenter, randomized, parallel-group, open-label trial. Patients were enrolled from February 2000 to February 2002 and followed up for 72 weeks. Four hundred twelve HIV-HCV coinfected patients with detectable serum HCV-RNA, abnormal liver histology, a CD4 cell count of at least 200 x 10(6)/L, and stable plasma HIV-RNA. Treatment with ribavirin 400 mg twice a day, orally, plus either peginterferon alfa-2b (1.5 microg/kg subcutaneous injection once a week) or standard interferon alfa-2b (3 million units of subcutaneous injection 3 times a week) for 48 weeks. Sustained virologic response, defined by undetectable serum HCV-RNA at week 72. More patients had sustained virologic responses in the peginterferon group than in the standard interferon group (27% vs 20%, P = .047). This difference between the treatments was found in patients with HCV genotype 1 or 4 infection (17% for peginterferon vs 6% for standard interferon, P = .006) but was not found in patients with HCV genotype 2, 3, or 5 (44% for peginterferon vs 43% for standard interferon, P = .88). Together, a decline in HCV-RNA of less than 2 log10 from baseline and detectable serum HCV-RNA at week 12 predicted 99% of treatment failures. Histologic activity diminished and fibrosis stabilized in virologic responders. The 2 regimens showed similar tolerability although dose modifications for clinical and biological events were more frequent with peginterferon. Eleven cases of pancreatitis or symptomatic hyperlactatemia were observed, all in patients receiving didanosine-containing antiretroviral regimens. In combination with ribavirin, treatment with peginterferon alfa-2b is more effective than standard interferon alfa-2b for HCV infection in HIV-infected patients.

Prediction of treatment outcome in patients with chronic hepatitis C: Significance of baseline parameters and viral dynamics during therapy

In patients with chronic hepatitis C virus (HCV) infection scheduled for a 48-week treatment period, premature discontinuation of treatment was previously recommended if HCV-RNA levels remained detectable at week 24 of therapy. Considering the number of side effects and treatment costs, measurement of initial viral decline during therapy may identify virologic nonresponse earlier than 24 weeks. We retrospectively analyzed 260 European patients treated with standard or pegylated interferon alfa (IFN-α) and ribavirin for 24 to 48 weeks. Early prediction of virologic response by HCV-RNA decline at weeks 4 and 12 (Versant Quantitative [branched DNA (bDNA) 3.0]; Bayer Diagnostics, Emeryville, CA; and Qualitative [transcription-mediated amplification (TMA)] HCV RNA assay; Bayer Diagnostics) as well as clinical, biochemical, virologic, and histologic baseline parameters were analyzed by logistic regression and receiver operating characteristic (ROC) curves. A viral load at treatment week 4 above 450,000 IU/mL and at week 12 above 30,000 IU/mL was 100% predictive for virologic nonresponse in all patients. From multivariate logistic regression analysis of all patients, independent predictors for sustained virologic response were: genotypes 2 and 3 ( P < .0001), a low baseline gamma-glutamyltransferase (GGT) level ( P < .0001), a high baseline alanine aminotransferase level ( P = .002), and a low baseline viral load ( P = .04). None of the latter 3 factors were predictive for sustained virologic response when analysis was restricted to the subgroup of genotypes 2– and 3–infected patients. In conclusion, virologic nonresponse can be predicted early at week 12 of treatment independent from the applied therapeutic regimen based on a cutoff level for HCV RNA of 30,000 IU/mL. This algorithm recognizes 53.7% of nonresponders previously identified at week 24 of treatment. (H EPATOLOGY 2003;37:600-609.)

Rapid HCV RNA decline after daily 18 MU interferon induction combined with ribavirin and amantadine in chronic HCV patients

Rapid HCV RNA decline after daily 18 MU interferon induction combined with ribavirin and amantadine in chronic HCV patients

Viral kinetics in patients with chronic hepatitis C treated with standard or peginterferon α2a

Covalent attachment of a 40-kilodalton polyethylene glycol moiety to interferon alpha2a (peginterferon alpha2a) results in sustained delivery and reduced clearance compared with standard interferon alpha2a. The aim of the study was to compare viral kinetics in patients treated with standard or peginterferon alpha2a. Patients with chronic hepatitis C were randomly assigned to receive either standard interferon alpha2a thrice weekly (n = 16) or 180 microg peginterferon alpha2a once weekly (n = 17) for 48 weeks. HCV RNA was quantitated before and frequently during treatment. The extent of the second-phase decline of HCV RNA, representing the degradation rate of infected cells during therapy for responding patients, was 0.02 +/- 0.03 day(-1) (HCV-1), 0.88 +/- 0.64 day(-1) (HCV non-1), 0.06 +/- 0.08 day(-1) (HCV-1), and 0.44 +/- 0.33 day(-1) (HCV non-1) in patients treated with standard or peginterferon alpha2a, respectively. The second-phase decline was low (<0.05 day(-1)) in most patients without a virological end-of-treatment response, and the second-phase decline was high (>0.25 day(-1)) in all patients with sustained virological response. The degradation rate of infected cells is HCV genotype dependent. Treatment with peginterferon alpha2a may reinforce the death rate of infected cells (particularly in HCV-1-infected patients) or stabilize the therapeutic effect on viral production. The second-phase decline of HCV RNA is predictive of virological sustained response.

HCVRNA increase during antiretroviral treatment (ART) induced HIVRNA rapid phase decline: An evidence of virus-virus interaction?

Background: Liver damage has been described in HIV+ patients with HCV coinfeetion undergoing combination ART. Aims and Methods. In order to assess the relationship between CD4, CD8,HIVRNA, HCVRNA (both measured by bDNA) and liver enzymes during combination antiretroviral treatment (ART), we measured these parameters in 12 HIVHCV coinfected patients on the day before, 3, 7, 14, 28, 56 and 84 days after starting: d4T and 3TC in all, Indinavir in 6 and Nevirapine in 6. Results: HIVRNA declined rapidly and significantly. CD4 raised slowly. CD8 cells and liver enzymes were stable. HCVRNA show a significant increase (7.55 +/at day 14 7.61 +/-at day 21 vs. 7.35 +/at baseline M+/SE log copies/mi p<0.05) independent from Indinavir use. Increase of HCVRNA levels was observed in 10/12 patients all showing a 2 log decrease of HIVRNA and was not related to liver damage. An ALT increase greater than 100% was observed in 4/12 patients all showing HCVRNA increase. Conclusions: The relationship between HCVRNA increase and decline of HIVRNA suggests interference between the two viruses. HCVRNA increase may elicit liver damage only in a minority of patients. Viral hepatitis: basic aspects

Quantification of the initial decline of serum hepatitis C virus RNA and response to interferon alfa.

Although several virus- and host-related predictive factors for the response to interferon alfa (IFN-alpha) have been defined in patients with chronic hepatitis C, no pretreatment parameter can definitely predict the response to antiviral treatment. Assessment of the initial response by quantification of serum hepatitis C virus RNA before and 4 weeks after initiation of therapy may be a clinically applicable and reliable parameter to predict long-term response. Therefore, the aims of the present study were to test the predictive value of a decline in HCV RNA of at least 3 log in the first 4 weeks of treatment (deltaHCV RNA) in patients treated with 3 x 10(6) units of recombinant IFN-alpha2a (rIFN-alpha2a) three times per week subcutaneously and to compare deltaHCV RNA with other established predictive factors, such as HCV genotype and pretreatment viremia. Serum HCV RNA was measured by a validated quantitative reverse transcription-polymerase chain reaction (RT-PCR). Geno/subtyping of HCV was performed by direct sequencing of the nonstructural (NS) 5B region of PCR-amplified isolates and subsequent phylogenetic analysis. Stable HCV RNA levels (deltaHCV RNA < or = 1 log) within the first 4 weeks of IFN-alpha treatment were present in 42 of 70 patients. A decline in HCV RNA levels between 1 to 3 log and more than 3 log was observed in 9 (13%) and 19 patients (27%), respectively. In 21 of 70 patients (30%), HCV RNA was not detectable at the end of 12 months' treatment. Three of 26 patients (11%) with a pretreatment viremia of < or = 10(6) copies/mL (all HCV subtype 3a) and 6 of 44 patients (14%) with a pretreatment viremia of > 10(6) copies/mL (HCV subtypes 1b, 2a, 2c, 3a [two patients], and 4) achieved a virological sustained response to interferon-alpha2a treatment. All patients with a virological sustained response had an initial deltaHCV RNA of more than 3 log. In a stepwise discriminant-function analysis, the initial deltaHCV RNA was confirmed as the strongest predictor of virological sustained response (P < .0001). In conclusion, the data of the present study suggest that IFN-alpha treatment can be terminated after 4 weeks in patients with a decrease in HCV RNA levels of less than 3 log, when apparent HCV eradication is considered the therapeutic target. The predictive value of deltaHCV RNA clearly exceeds the significance of HCV genotype and pretreatment viremia as predictors of successful IFN-alpha treatment.

A quantitative HCV-PCR test for routine diagnostics

The aim of this study was to develop a reliable and simple method for hepatitis C virus (HCV)-PCR using standard, automated laboratory equipment. HCV-RNA was extracted from serum and amplified in a single PCR with an internal standard. The PCR product was detected using fluoroimmunoassay. Quantification was based on external and internal standards. Linearity was observed over a wide range (10-107 geq). Mean inter and intra serial coefficients of variation were 35% and 23%, respectively. The limit of quantification was 1000 geq/ml based on intra and inter serial variations, while levels of 110 geq/ml were always detectable. Lower concentrations were intermittently positive. The ability to separate HCV-signals in healthy and infected persons was good, based on the distribution of HCV-signals from 353 random blood donors and 191 patient samples. To illustrate the applicability of the test, HCV-RNA quantification was performed in 11 patients during treatment with interferon alpha-2b. Ten of 11 patients showed a decline in HCV-RNA within the first few weeks of treatment. After four weeks most patients were still HCV-RNA positive but below the limit of quantification. The present method for quantification of HCV-RNA was shown to have sensitivity at the level of nested PCR techniques. Until now HCV-PCR has been complicated, time-consuming and costly, and therefore not suitable for routine diagnostics. The PCR method described here is easy to perform, fast and cost-effective.