Molecular Detection of Hepatitis C Virus amongst Patients in Five Selected Hospitals in Niger State, Nigeria

2019 ◽  
Vol 2 (1) ◽  
pp. 60-69
Author(s):  
M U Iduh ◽  
F A Kuta ◽  
M E Abalaka ◽  
K O Shitu
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Direct Sequencing ◽  
Rna Extraction ◽  
Public Health Problem ◽  
Developed Countries ◽  
Response To Treatment ◽  
Hcv Rna ◽  
Major Public Health Problem ◽  
Polymerase Chain

Hepatitis C Virus (HCV) is a major public health problem in developing and developed countries worldwide. It is responsible for liver diseases and hepatocellular carcinoma in chronically-infected patients. This study therefore aimed to identify the strain of HCV among HCV seropositive subjects in Niger State. A total of 44 HCV seropositive blood samples which consisted of 27 males and 17 females were analyzed (after Viral RNA extraction) for the presence of HCV-RNA by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Nine (20.5%) of the samples were positive for HCV RNA. HCV-RNA positive samples were genotyped by direct sequencing at 5’UTR region genomes; sequences were aligned on MEGA 6.0 and confirmed by phylogenetic analysis. HCV genotype 1b was the only one distributed among the participants. The findings are relevant as predictors for using antiviral therapy in this population because the response to treatment varies according to the genotype.

scholarly journals Molecular Detection of Hepatitis C Virus amongst Patients in Five Selected Hospitals in Niger State, Nigeria

2019 ◽  
Vol 2 (1) ◽  
pp. 60-69
Author(s):  
M U Iduh ◽  
F A Kuta ◽  
M E Abalaka ◽  
K O Shitu
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Direct Sequencing ◽  
Rna Extraction ◽  
Public Health Problem ◽  
Developed Countries ◽  
Response To Treatment ◽  
Hcv Rna ◽  
Major Public Health Problem ◽  
Polymerase Chain

Hepatitis C Virus (HCV) is a major public health problem in developing and developed countries worldwide. It is responsible for liver diseases and hepatocellular carcinoma in chronically-infected patients. This study therefore aimed to identify the strain of HCV among HCV seropositive subjects in Niger State. A total of 44 HCV seropositive blood samples which consisted of 27 males and 17 females were analyzed (after Viral RNA extraction) for the presence of HCV-RNA by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Nine (20.5%) of the samples were positive for HCV RNA. HCV-RNA positive samples were genotyped by direct sequencing at 5’UTR region genomes; sequences were aligned on MEGA 6.0 and confirmed by phylogenetic analysis. HCV genotype 1b was the only one distributed among the participants. The findings are relevant as predictors for using antiviral therapy in this population because the response to treatment varies according to the genotype.

scholarly journals Molecular Detection of Hepatitis C Virus amongst Patients in Five Selected Hospitals in Niger State, Nigeria

2019 ◽  
Vol 2 (1) ◽  
pp. 60-69
Author(s):  
M U Iduh ◽  
F A Kuta ◽  
M E Abalaka ◽  
K O Shitu
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Direct Sequencing ◽  
Rna Extraction ◽  
Public Health Problem ◽  
Developed Countries ◽  
Response To Treatment ◽  
Hcv Rna ◽  
Major Public Health Problem ◽  
Polymerase Chain

Hepatitis C Virus (HCV) is a major public health problem in developing and developed countries worldwide. It is responsible for liver diseases and hepatocellular carcinoma in chronically-infected patients. This study therefore aimed to identify the strain of HCV among HCV seropositive subjects in Niger State. A total of 44 HCV seropositive blood samples which consisted of 27 males and 17 females were analyzed (after Viral RNA extraction) for the presence of HCV-RNA by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Nine (20.5%) of the samples were positive for HCV RNA. HCV-RNA positive samples were genotyped by direct sequencing at 5’UTR region genomes; sequences were aligned on MEGA 6.0 and confirmed by phylogenetic analysis. HCV genotype 1b was the only one distributed among the participants. The findings are relevant as predictors for using antiviral therapy in this population because the response to treatment varies according to the genotype.

scholarly journals Influence of OASL gene polymorphisms on host response to interferon therapy in chronic hepatitis C virus patients

2017 ◽  
Vol 1 (2) ◽  
pp. 117-125
Author(s):  
Sanja Kiprijanovska ◽  
Emilija Sukarova Stefanovska ◽  
Predrag Noveski ◽  
Viktorija Chaloska Ivanova ◽  
Dijana Plaseska-Karanfilska
Keyword(s):  
Chronic Hepatitis ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Chronic Hepatitis C ◽  
Host Response ◽  
Standard Treatment ◽  
Public Health Problem ◽  
Hcv Rna ◽  
Major Public Health Problem ◽  
Oligoadenylate Synthetase

Abstract Hepatitis C virus (HCV) infection becomes a major public health problem and leading cause of chronic liver disease and liver failure. Pegylated interferon-alfa and ribavirin are currently the standard treatment for chronic hepatitis C (CHC). It is indicated that genes that trace the interferon signaling could be associated with the host response to the therapy. In order to investigate the influence of these genes on host related response, we have analyzed seven single nucleotide polymorphisms (rs59248852, rs74390571, rs12811390, rs1169279, rs3213545, rs1083129 and rs2859398) in 2-5-Oligoadenylate- Synthetase-Like (OASL) gene in CHC cases from Republic of Macedonia. A simple and easy to use SNaPshot method was developed. A cohort of 100 HCV RNA positive patients - non responders and 109 patients with achieved virological response after the standard treatment were included in this study. We have found significant association in five of the seven studied SNP` s: rs59248852 [p = 6.5x10-31, OR=55.7 (20.0-155.3)]; rs12811390 [p = 2.2x10-11, OR=4.3 (2.3-6.7)]; rs2859398 [p=1.34x10-9, OR=3.4 (2.2-5.0)]; rs74390571 [p=4.3x10-7, OR=2.9 (1.9-4.4)], and rs1083129 [p=0.0139, OR=2.0 (1.1-3.5)]. The results from this study can be used as a predictive factor of future patient’s selection for the standard therapy, having an important cost benefit for the health insurance system.

Detection of active hepatitis C virus and hepatitis G virus/GB virus C replication in bone marrow in human subjects

Blood ◽  
2000 ◽  
Vol 95 (12) ◽  
pp. 3986-3989 ◽  
Author(s):  
Marek Radkowski ◽  
Joanna Kubicka ◽  
Elzbieta Kisiel ◽  
Janusz Cianciara ◽  
Marek Nowicki ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Human Subjects ◽  
Hcv Rna ◽  
Serum Samples ◽  
Hepatitis G Virus ◽  
Polymerase Chain ◽  
Hepatitis G ◽  
Virus C

Abstract We have analyzed the presence of hepatitis C virus (HCV) and hepatitis G virus (HGV) sequences in bone marrow and serum samples from 48 patients of a hematologic outpatient clinic. HCV RNA was detected in 18 (38%) and 15 (31%) and HGV RNA was detected in 6 (13%) and 9 (19%) of serum and bone marrow samples, respectively. In 3 patients, HGV RNA was detectable in bone marrow but not in the serum; 2 of these patients were negative for the presence of specific antibodies. Using a highly strand-specific Tth-based reverse transcriptase-polymerase chain reaction (RT-PCR), the presence of HCV RNA and HGV RNA negative strand was demonstrated in 4 and 5 bone marrow samples, respectively. Our study shows that HCV and HGV can replicate in bone marrow; in the case of HGV, analysis of serum may underestimate the true prevalence of infection.

Detection of active hepatitis C virus and hepatitis G virus/GB virus C replication in bone marrow in human subjects

Blood ◽  
2000 ◽  
Vol 95 (12) ◽  
pp. 3986-3989 ◽  
Author(s):  
Marek Radkowski ◽  
Joanna Kubicka ◽  
Elzbieta Kisiel ◽  
Janusz Cianciara ◽  
Marek Nowicki ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Human Subjects ◽  
Hcv Rna ◽  
Serum Samples ◽  
Hepatitis G Virus ◽  
Polymerase Chain ◽  
Hepatitis G ◽  
Virus C

We have analyzed the presence of hepatitis C virus (HCV) and hepatitis G virus (HGV) sequences in bone marrow and serum samples from 48 patients of a hematologic outpatient clinic. HCV RNA was detected in 18 (38%) and 15 (31%) and HGV RNA was detected in 6 (13%) and 9 (19%) of serum and bone marrow samples, respectively. In 3 patients, HGV RNA was detectable in bone marrow but not in the serum; 2 of these patients were negative for the presence of specific antibodies. Using a highly strand-specific Tth-based reverse transcriptase-polymerase chain reaction (RT-PCR), the presence of HCV RNA and HGV RNA negative strand was demonstrated in 4 and 5 bone marrow samples, respectively. Our study shows that HCV and HGV can replicate in bone marrow; in the case of HGV, analysis of serum may underestimate the true prevalence of infection.

scholarly journals Automated Specific Capture of Hepatitis C Virus RNA with Probes and Paramagnetic Particle Separation

2000 ◽  
Vol 38 (1) ◽  
pp. 18-21
Author(s):  
Hayato Miyachi ◽  
Atsuko Masukawa ◽  
Toshio Ohshima ◽  
Toru Hirose ◽  
Chaka Impraim ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Hemodialysis Patient ◽  
Rna Extraction ◽  
Magnetic Bead ◽  
Inhibitory Effect ◽  
Hcv Rna ◽  
Specimen Preparation ◽  
Manual Method ◽  
Fluid Separation

ABSTRACT We developed and evaluated a prototype automated specimen preparation instrument for the specific capture of hepatitis C virus (HCV) RNA with probes and magnetic bead-fluid separation. HCV RNA was isolated from serum by lysis of virus particles with a chaotropic agent, followed by hybridization of the RNA with biotinylated probes and capture of the hybridized RNA with streptavidin-coated paramagnetic particles. After washing of the hybrid-particle complexes to remove nonspecifically bound materials, the particles were resuspended in a specimen diluent and were then ready for amplification and detection with a fully automated PCR system (COBAS AMPLICOR; Roche Diagnostic Systems). The analytical sensitivity in the dilution series was 33 copies per ml or greater. Comparison of the test results with those obtained by a manual method based on organic extraction and precipitation of RNA (SepaGene RV-R; Sanko Junyaku Co., Ltd.) showed 93% (49 of 53 samples) sensitivity and 100% (12 of 12 samples) specificity. There was 94% overall agreement between results. When RNA was extracted by the manual method from serum containing 10 3 or 10 5 copies of HCV per ml in the presence of heparin, there was an inhibitory effect on detection of both HCV RNA and the internal control. In contrast, when RNA was extracted from the serum by the automated method, there was no inhibitory effect. This inhibitory effect of heparin on the manual method was also observed for a series of serum specimens from a hemodialysis patient, but the inhibitory effect was eliminated by the automated specimen preparation method. In summary, a fully automated RNA extraction system for PCR detection of HCV RNA by use of specific capture with probes and magnetic bead-fluid separation was shown to have performance similar to that of the conventional manual method. In addition, it successfully eliminated the inhibitory effect of the heparin in the serum and permitted the detection of HCV RNA in serum samples from a hemodialysis patient. The prototype automated RNA extraction system is suitable as a totally automated system, starting with RNA extraction to detection of HCV, if it was combined with the fully automated COBAS AMPLICOR PCR system.

scholarly journals Early viral kinetics and response to treatment in a hepatitis C virus genotype 5 infected patient

2011 ◽  
Vol 1 (2) ◽  
pp. 28
Author(s):  
Emanuele Durante-Mangoni ◽  
Domenico Iossa ◽  
Umberto Malgeri
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Antiviral Treatment ◽  
Pegylated Interferon ◽  
Response To Treatment ◽  
Core Antigen ◽  
Hcv Rna ◽  
Virus Genotype ◽  
Viral Kinetics ◽  
Genotype 5

Genotype 5 hepatitis C has been poorly studied despite its worldwide spread. We have analyzed the early kinetics of genotype 5 hepatitis C virus RNA during pegylated interferon/ribavirin treatment in a 59-year-old man with active liver necroinflammatory changes and advanced liver fibrosis. The patient had a high viral load but a small serum level of hepatitis C core antigen. On combination antiviral treatment with pegylated-interferon alpha 2a, 180 ?g/week, and ribavirin, 1200 mg/day, the patient experienced an impressive reduction in serum HCV RNA as early as day 2 of treatment and eventually became a sustained virological responder. Our viral kinetics data support previous clinical studies showing HCV genotype 5 could be as intrinsically sensitive to interferon as HCV genotypes 2 and 3.

scholarly journals Updating the Signal-to-cutoff Level to Reduce Anti-hepatitis C Virus False Positivity

2021 ◽  
Vol 14 (10) ◽  
Author(s):  
Servet Oztürk ◽  
Canan Ağalar
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Area Under The Curve ◽  
Hcv Rna ◽  
Hepatitis C Infection ◽  
False Positivity ◽  
History Of ◽  
Training And Research ◽  
Polymerase Chain ◽  
The Cost

Background: Anti-hepatitis C virus (anti-HCV) is the only screening test being used in the diagnosis of hepatitis C. In this study, we examined anti-HCV positivity rates in our hospital. Objectives: The aim of administering the anti-HCV test was to distinguish patients with hepatitis C infection from false positivity in patients with reactive results. Methods: The anti-HCV tests were performed at Fatih Sultan Mehmet Training and Research Hospital in Istanbul, Turkey, between January 1, 2015 and December 31, 2019. The patients were evaluated retrospectively in terms of age, gender, anti-HCV titer, the clinic for which the examination was requested, the reason for the examination, and the history of hepatitis C. Results: In this study, 511 patients who had two negative polymerase chain reaction (PCR) results were evaluated as false positive cases and enrolled. The cut-off value was found to be 7.5 IU/ml, with the highest sensitivity of 94.4% and specificity of 94.5% (area under the curve [AUC]: 0.982). The lowest anti-HCV titer (5.2) was from patients without acute hepatitis, who were HCV-RNA positive and diagnosed with chronic hepatitis C. Conclusions: It may be more appropriate to report anti-HCV cut-off value of 0 - 5 as negative, 5 - 7.5 as borderline, and > 7.5 as positive. Working with a more acceptable cut-off level with a greater number of tests can help identify patients with asymptomatic HCV infection. Also, it can possibly reduce the cost due to a decrease in the number of PCR tests administered.

scholarly journals Fully Automated Detection of Hepatitis C Virus RNA in Serum and Whole-Blood Samples

2002 ◽  
Vol 9 (6) ◽  
pp. 1385-1388 ◽  
Author(s):  
Harald H. Kessler ◽  
Alexandra M. K. Clarici ◽  
Evelyn Stelzl ◽  
Gerhard Mühlbauer ◽  
Elisabeth Daghofer ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Whole Blood ◽  
Rna Extraction ◽  
Hcv Rna ◽  
Molecular Assay ◽  
Blood Samples ◽  
Molecular Assays ◽  
Roche Molecular Diagnostics ◽  
Whole Blood Samples

ABSTRACT In this study, we established a fully automated molecular assay for qualitative detection of hepatitis C virus (HCV) in serum and whole-blood samples and compared it with conventional molecular assays, including manual HCV RNA extraction protocols. Whole-blood samples were collected from patients with and without chronic HCV infection in EDTA tubes and nucleic acid stabilization tubes (NASTs). Prior to HCV RNA extraction, the HCV Internal Control (IC), derived from the COBAS AMPLICOR HCV test, version 2.0 (Roche Molecular Diagnostics), was added. The new assay was based on an automated extraction protocol on the MagNA Pure LC instrument (Roche Applied Science), followed by automated reverse transcription, amplification, hybridization, and detection on the Cobas Amplicor analyzer (Roche Molecular Diagnostics). The detection limit of the new assay was found to be similar to those of conventional molecular assays. In clinical samples, 100% agreement between the new assay and conventional methods was observed. The introduced amount of IC was detected in 45 of 45 serum samples, 41 of 45 EDTA tube whole-blood samples, and 43 of 45 NAST whole-blood samples. Retesting led to more frequent IC detection. The fully automated molecular assay was found to be suitable for detection of HCV RNA in different kinds of sample materials. It may be recommended for use in the high-throughput routine molecular diagnostic laboratory.

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