Improved diagnosis of chronic hepatitis C virus infection by detection of antibody to multiple epitopes: Confirmation by antibody to synthetic oligopeptides

Brown, David J.; Powell, Lee; Morris, A.; Rassam, Suha; Sherlock, Sheila; McIntyre, Neil; Zuckerman, Arie J.; Dusheiko, Geoffrey

doi:10.1002/jmv.1890380303

Cited by 14 publications

(7 citation statements)

References 21 publications

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“…The presence of HCV RNA in serum was determined by reverse-transcription nested polymerase chain reaction (RT-PCR) using primers from the 5Ј NCR region, as described previously [Brown et al, 1992] .…”

Section: Hcv Rna Detectionmentioning

confidence: 99%

GB virus C/hepatitis G virus infection in KwaZulu Natal, South Africa

et al. 1999

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Sera from 70 patients on maintenance haemodialysis, 98 patients with chronic liver disease, and 232 volunteer blood donors in the province of KwaZulu Natal, South Africa, were screened for GB virus/hepatitis G virus (GBV-C/HGV) RNA and anti-E2 by reverse transcription-polymerase chain reaction (RT-PCR) and by an enzyme-linked immunosorbent assay (ELISA), respectively. GBV-C/HGV RNA was detected in 17/70 (24.3%) haemodialysis patients, 12/98 (12.2%) patients with chronic liver disease, and 44/232 (18.9%) blood donors (Africans [29/76; 38.2%]; Asians [2/52; 3.8%]; Whites [11/49; 22.4%], and "Coloureds" [persons of mixed origin; 2/55; 3.6%]). Overall (anti-E2 and/or RNA) 43.9% (43/98) of patients with chronic liver disease, 47.1% (33/70) of haemodialysis patients, and 31.9% (74/232) of blood donors (Africans [44/76; 5.9%]; Asians [5/52; 9.6%]; Whites [15/49; 30.6%], and Coloureds [9/54; 16.6%]) were exposed to GBV-C/HGV infection. There was a significant difference in the prevalence of GBV-C/HGV infection (RNA and/or anti-E2) between African blood donors and the other racial groups (P < .001), and between blood donors and haemodialysis patients (P = .02) and patients with chronic liver disease (P = .04). Anti-E2 antibodies and GBV-C/HGV RNA were almost mutually exclusive. GBV-C/HGV-infected haemodialysis patients received more transfusions (P = .03) than noninfected patients. There was no significant difference in liver biochemistry between GBV-C/HGV-infected and noninfected patients and between blood donors in each of the four racial groups. The high prevalence of GBV-C/HGV infection in blood donors and chronic liver disease patients, and the lack of elevated liver enzymes and clinical hepatitis in blood donors and haemodialysis patients, suggest that GBV-C/HGV may not be associated with liver disease.

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Section: Hcv Rna Detectionmentioning

confidence: 99%

GB virus C/hepatitis G virus infection in KwaZulu Natal, South Africa

et al. 1999

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“…Evidence of HCV infection is found in 60 to 100% of patients with chronic hepatitis when other causes are excluded and EIA-2 screening tests are used (38,118,148,260,311). Higher rates of HCV infection are observed in Japan and Mediterranean countries, and lower rates are seen in the United States and the United Kingdom.…”

Section: Chronic Hepatitismentioning

confidence: 99%

Hepatitis C: progress and problems

Cuthbert

1994

Clin Microbiol Rev

153

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The hepatitis C virus (HCV), a single-stranded RNA virus, is the major cause of posttransfusion hepatitis. HCV isolates differ in nucleotide and amino acid sequences. Nucleotide changes are concentrated in hypervariable regions and may be related to immune selection. In most immunocompetent persons, HCV infection is diagnosed serologically, using antigens from conserved regions. Amplification of RNA may be necessary to detect infection in immunosuppressed patients. Transmission by known parenteral routes is frequent; other means of spread are less common and may represent inapparent, percutaneous dissemination. Infection can lead to classical acute hepatitis, but most infected persons have no history of acute disease. Once infected, most individuals apparently remain carriers of the virus, with varying degrees of hepatocyte damage and fibrosis ensuing. Chronic hepatitis may lead to cirrhosis and hepatocellular carcinoma. However, disease progression varies widely, from less than 2 years to cirrhosis in some patients to more than 30 years with only chronic hepatitis in others. Determinants important in deciding outcome are unknown. Alpha interferon, which results in sustained remission in selected patients, is the only available therapy. Long-term benefits from such therapy have not been demonstrated. Prevention of HCV infection by vaccination is likely to be challenging if ongoing viral mutation results in escape from neutralization and clearance.

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“…Details of the PCR method were as described by Brown et al [1992]. In short, 5 pl of the cDNA solution was added to a first round PCR mix containing 50 pM of each of the outer primers, 0.2 mM dNTPs, 10 p1 of l o x Taq polymerase buffer, 1.5 mM MgCl,, and 2.5 U of Taq DNA polymerase (Gibco) in a total reaction volume of 100 pl overlaid with a few drops of mineral oil (Sigma).…”

Section: Pcrmentioning

confidence: 99%

“…Hepatitis C virus (HCV) was first identified [Choo et al, 19891 and shown to be a small enveloped RNA virus 0 1994 WILEY-LISS, INC. [Houghton et al, 19911 using molecular biological techniques. The polymerase chain reaction (PCR) has been used by many investigators to detect HCV-RNA in serum [Garson et al, 1990;Brown et al, 1992;Kato et al, 1993;Yun et al, 19931 or liver tissue LWeiner et al, 1990;Shieh et al, 1991;Sallie et al, 19921 of patients with hepatitis C. In contrast with the delayed appearance of anti-HCV (30-90 days postinfection), detection of serum HCV-RNA by PCR is a n early event, occurring as soon as 5-10 days after infection [Trepo et al, 19931. A major advantage of the PCR technique is its extreme sensitivity that has led to the detection of a low copy number of virus in individuals who were negative for conventional viral markers [Bonino et al, 19921. While allowing a direct and highly sensitive identification of viral genome, PCR however, has certain limitations imposed, for example, by possible contamination problems [Brechot, 19931. Recently, 31 laboratories cooperated in a quality control study for detecting HCV-RNA in serum by PCR; false-negative as well as falsepositive results were reported [Zaaijer et al, 19931. Bresters et al [1992] reported a 100% correlation between HCV cDNA-PCR results from plasma, freshfrozen and formalin-fixed, paraffin embedded (FFPE) liver tissue of patients with chronic non-A, non-B hepatitis who had not been treated with antiviral agents.…”

Section: Introductionmentioning

confidence: 99%

Hepatitis C virus‐polymerase chain reaction of routinely processed liver biopsies

El-Batanony

Savage

Jacobs

et al. 1994

Journal of Medical Virology

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The aim of the study was to evaluate the specificity and sensitivity of detection of hepatitis C virus (HCV)-RNA in formalin-fixed paraffin-embedded (FFPE) liver biopsies by polymerase chain reaction (PCR). Routinely processed FFPE diagnostic needle liver biopsies as well as stored serum samples from 43 patients with liver disease were tested for HCV-RNA by reverse transcription-nested PCR using the same sets of primers and following strict anticontamination measures. Twenty-nine cases were positive and 14 were negative for serum HCV-RNA. Tissue HCV-RNA was detected in 17 out of the 29 serum HCV-RNA-positive cases but not in any of the 14 serum HCV-RNA-negative cases. Compared to serum-PCR, tissue-PCR was 100% specific, 58.6% sensitive, and 72% efficient. HCV-RNA was detected more frequently in biopsies stored for less than 1 year, than in those stored for more than 1 year (P = 0.046). In biopsies stored for up to 1 year detection of HCV-RNA by PCR was 81.8% sensitive and 90.9% efficient. Short (< 0.5 cm) liver biopsies were as sufficient for nucleic acid extraction and amplification as long (> 0.5 cm) ones. It is concluded that following strict anticontamination measures, HCV-RNA detection by PCR in routinely fixed, processed, and stored diagnostic liver biopsies provides a valuable adjunct to diagnosis of HCV infection. In this study, this option was free from contamination problems, even though routine batch histological processing schedules were used.

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Improved diagnosis of chronic hepatitis C virus infection by detection of antibody to multiple epitopes: Confirmation by antibody to synthetic oligopeptides

Cited by 14 publications

References 21 publications

GB virus C/hepatitis G virus infection in KwaZulu Natal, South Africa

GB virus C/hepatitis G virus infection in KwaZulu Natal, South Africa

Hepatitis C: progress and problems

Hepatitis C virus‐polymerase chain reaction of routinely processed liver biopsies

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