Enhanced detection by PCR of hepatitis C virus RNA

Garson, Jeremy A.; Ring, Christopher J.; Tuke, Philip W.; Tedder, Richard S.

doi:10.1016/0140-6736(90)92384-t

Cited by 199 publications

(91 citation statements)

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“…These patients were seronegative for HBsAg, anti-HBc and anti-HCV antibodies as determined with commercially available kits (Abbott Laboratories, Chicago, IL, USA). Acute cases also tested negative for anti-HAV IgM, anti-HBc IgM, anti-HEV antibodies (Abbott) and HCV-RNA by RT-PCR using previously described primers (15,16) after guanidine isothiocyanate extraction (17).…”

Section: Patientsmentioning

confidence: 99%

Detection of human parvovirus B19 in a patient with hepatitis

Pinho

Alves

Vieira

et al. 2001

Braz J Med Biol Res

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Parvovirus B19 has been associated by some investigators with cases of severe hepatitis. The aim of the present study was to determine the presence of active parvovirus B19 infection among 129 Brazilian patients with non-A-E hepatitis. The patients were assayed for antibodies against parvovirus B19, IgM class, by ELISA. In IgM-positive cases, parvovirus B19 DNA was assayed by PCR in serum and liver tissue and parvovirus VP1 antigen in liver tissue was assayed by immunohistochemistry. Antibodies against parvovirus B19, IgM class, were detected in 3 (2.3%) of 129 patients with non-A-E hepatitis. Previous surgery and blood transfusions were reported by these 3 patients. One patient was a 56-year-old female with severe hepatitis, with antimitochondrial antibody seropositivity and submassive necrosis at liver biopsy, who responded to corticosteroid therapy. Strong evidence for active parvovirus B19 infection was found in this patient, with parvovirus B19 DNA being detected by PCR in liver tissue. Furthermore, parvovirus VP1 antigen was also detected in liver tissue by immunohistochemistry. The other two IgM-positive patients were chronic hepatitis cases, but active infection was not proven, since neither viral DNA nor antigen were detected in their liver tissues. This and other reports suggest a possible relation between parvovirus B19 infection and some cases of hepatitis.

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Section: Patientsmentioning

confidence: 99%

Detection of human parvovirus B19 in a patient with hepatitis

Pinho

Alves

Vieira

et al. 2001

Braz J Med Biol Res

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“…Using PCR assay, the data obtained show that viremia can be detected within only a few days of exposure to the virus and many weeks before elevation of transaminases and antibody titers. 28,29,30 Furthermore, PCR offers a possibility to monitor anti-viral treatment for chronic HCV infection 31 and recent evidence indicates that quantitative PCR is important as the response to interferon therapy depends on the viral load. 32 Also, PCR may be useful for diagnosing vertical transmission of HCV from chronically infected mothers to their offspring.…”

Section: Detection Of Viral Rnamentioning

confidence: 99%

Hepatitis C Virus (HCV) Infection in Saudi Arabia: A Review

Al-Faleh

Ramia

1997

Ann Saudi Med

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It is well established now that HCV is the major etiological agent of parenterally transmitted non-A, non-B hepatitis (PT-NANBH), [1][2][3] and that it has a worldwide distribution. Studies on HCV infection have led to three striking observations. First, there is a high frequency of chronic infection in a significant number of infected individuals. It is estimated that at least 50% of HCV infections lead to chronic liver disease, including chronic active hepatitis with or without concurrent cirrhosis. [4][5][6] Second, HCV has been implicated as one of the major causative agents of primary hepatocellular carcinoma (HCC) in Japan 7 , Saudi Arabia, 8 and other parts of the world. [9][10] Third, approximately 45% of HCV cases have no obvious risk factors, including parenteral exposure, 4-6 leaving unanswered the question of virus transmission via as yet unidentified routes of exposure. The aim of this article is to review the literature about the extent of HCV infection in Saudi Arabia and see what can be concluded from the studies conducted so far. However, since there have been dramatic advances in the serologic diagnosis of HCV during the past six to seven years, we would first like to briefly review those serologic tests used and their reliability in diagnosing HCV infection. Diagnosis of HCV Infection Anti-HCV enzyme immunoassay (EIA) testsHCV infection, once a diagnosis of "exclusion" based on the absence of markers of acute infection with hepatitis A virus (HAV) or hepatitis B virus (HBV) can now be specifically diagnosed by using serodiagnostic assays for virus-specific antibodies.11 A first-generation anti-HCV EIA test was developed by using recombinant c100-3, derived from the NS3/NS4 region of the genome of HCV, as antigen (Figure 1). The test was used widely and although important data was generated, the test suffered from major drawbacks. These included failure to discover all patients with HCV infection, 12 long "window-phase" before seroconversion (could be up to 12 months), 13 in addition to a high rate of false-positive reactions. [14][15][16][17][18][19][20] To circumvent the drawbacks of the anti-HCV c100-3 test, recombinant or synthetic antigens derived from other regions of the HCV genome were included in the test and this is what is referred to as second-generation anti-HCV EIA tests. The additional antigens c22 and c33 were derived from two conserved regions: the structural region (core) and NS3 region, respectively (Figure l). 21,22 In patients with posttransfusion NANBH, the seroconversion rate improved from 54% with the first-generation to 82% with a second-generation test and the time lag to seroconversion decreased from a mean of 6.1 weeks after the onset of hepatitis with the first-generation test to a mean of 2.3 weeks with the second-generation test.23 Thus, the second-generation test reduces the "window-phase" to seroconversion and increases the sensitivity in diagnosing HCV infection, with a dramatic reduction in the number of false-positive reactions seen with the first-generation ...

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“…The polymerase chain reaction (PCR) primers used for amplification of the NS5B (17), E1 (18), and 5'UTR (3,19) regions are given in Table 1. For the amplification of NS5B, semi-nested PCR was used, and for the amplification of the E1 and 5'UTR regions, nested PCR was used.…”

Section: Amplification Of Hcv Ns5b E1 and 5'utr Regionsmentioning

confidence: 99%

Hepatitis C virus genotype distribution in Turkey remains unchanged after a decade: Performance of phylogenetic analysis of the NS5B, E1, and 5’UTR regions in genotyping efficiency

Alagöz

Karataylı²,

Karataylı³

et al. 2014

Turk J Gastroenterol

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Background/Aims: Hepatitis C virus (HCV) genotyping has a considerable effect on therapy. The aim was to determine the change in prevalence of HCV genotypes in Turkey during the last decade and to compare the performance of DNA sequencing of different targets in the HCV genome (NS5B, E1, and 5'UTR). Materials and Methods: Five hundred HCV RNA-positive patients (226 males, 274 females) were included in the study. The NS5B, E1, and 5'UTR regions of the HCV genome were amplified by polymerase chain reaction (PCR) in patients where possible. Amplified PCR products were sequenced directly, and phylogenetic analysis was performed. A commonly used database, namely www.hcv.lanl.gov, was also used to determine the genotypes. Results: Phylogenetic analysis of the NS5B, E1, and 5'UTR regions showed that 1b was the most frequent genotype, with percentages of 92.5%, 93.5%, and 87.7%, respectively. Genotype 1a was the second most prevalent genotype, with ratios of 6.7%, 5.6%, and 6.6%, whereas genotype 2a was detected in proportions of 0.4%, 0.2%, and 0.8%, respectively. Genotype 5 or 6 was not detected among patients. The phylogenetic analysis showed discordant results with 18 patients' genotypes for different targets. The phylogenetic analysis showed similar results with the hcv.lanl. gov database for the E1 and NS5B sequences. Conclusion: There has been no change in genotyping profiles of Turkey during the last decade, representing 1b as the most prevalent subtype, followed by 1a. Phylogenetic analysis of HCV indicated high performance compared with the hcv.lanl.gov database when sequences of E1 and NS5B regions were analyzed.

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Enhanced detection by PCR of hepatitis C virus RNA

Cited by 199 publications

References 1 publication

Detection of human parvovirus B19 in a patient with hepatitis

Detection of human parvovirus B19 in a patient with hepatitis

Hepatitis C Virus (HCV) Infection in Saudi Arabia: A Review

Hepatitis C virus genotype distribution in Turkey remains unchanged after a decade: Performance of phylogenetic analysis of the NS5B, E1, and 5’UTR regions in genotyping efficiency

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