scholarly journals Detection and Quantification of HCV-RNA by RT-PCR among Anti-HCV Positive Patients

2020 ◽  
Vol 15 (1) ◽  
pp. 84-86
Author(s):  
Wasila Rahman ◽  
Md Rahimgir ◽  
Arif Ahmed Khan ◽  
Maj Suman Khisa ◽  
Rahima Akter ◽  
...  
Keyword(s):  
Hepatitis C ◽  
Real Time ◽  
Armed Forces ◽  
Income Group ◽  
Military Hospital ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hcv Rna ◽  
High Income Group ◽  
Rt Pcr ◽  
Group Family

Introduction: The most common contemporary strategy to diagnose chronic hepatitis C virus (HCV) infection consists of initial screening with an HCV enzyme immunoassay (EIA) antibody test followed by supplemental testing of positive screening tests with a quantitative HCV RNA assay to confirm the positive EIA and to determine whether they have active or resolved hepatitis C infection. Objectives: To detect and quantify HCV-RNA by real-time polymerase chain reaction (real-time PCR) among anti-HCV positive patients and to identify the socio demographic factors among these patients. Materials and Methods: This was a descriptive type of cross-sectional study which was conducted in Combined Military Hospital and Armed Forces Institute of Pathology, Dhaka cantonment. A total of 108 anti-HCV positive patients by enzyme-linked immunosorbent assay (ELISA), who were clinically suspected and advised for anti-HCV test, were selected randomly for the study and subjected to do HCV-RNA analysis during the period of October 2016 to September 2017. Results: Out of 108 anti-HCV positive patients by ELISA, HCV-RNA was detected in 72 (66.7%) cases with mean value of HCV RNA quantification was 2013323.95±2695207.41 (IU/ ml). Majority of anti-HCV positive patients (29.6%) belonged to 51-60 years age group with male predominance (58.33%). It was observed that 43.52% patients came from middle income group family, 31.48% came from poor and 25.0% came from high income group family. Risk factor for HCV infected population was found maximum in dialysis patients (47.37%), followed by blood transfusion (13.89%), Injecting drug User (IDUs) (12.04%), surgery & intervention (9.26%) and sexual transmission (1.85%). Mean alanine aminotransferase (ALT) was found 67.30±44.99 U/L among HCV-RNA detected patients (p< 0.05). Conclusion: The quantification of HCV RNA by RT-PCR will be helpful to rationalize the treatment, enhance antiviral responses and mitigate mortalities of HCV infected patients. Journal of Armed Forces Medical College Bangladesh Vol.15 (1) 2019: 84-86

scholarly journals Monitoring Current-Season Spread of Potato virus Y in Potato Fields Using ELISA and Real-Time RT-PCR

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 641-644 ◽  
Author(s):  
Manphool S. Fageria ◽  
Mathuresh Singh ◽  
Upeksha Nanayakkara ◽  
Yvan Pelletier ◽  
Xianzhou Nie ◽  
...  
Keyword(s):  
Real Time ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Current Season ◽  
Seed Market ◽  
Polymerase Chain ◽  
Time Of Harvest

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.

scholarly journals Demonstration of Hepatitis C Virus RNA withIn SituHybridization Employing a Locked Nucleic Acid Probe in Humanized Liver of Infected Chimeric Mice and in Needle-Biopsied Human Liver

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Kazuya Shiogama ◽  
Ken-ichi Inada ◽  
Michinori Kohara ◽  
Hidemi Teramoto ◽  
Yasuyoshi Mizutani ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Nucleic Acid ◽  
Oligonucleotide Probe ◽  
High Sensitivity ◽  
Locked Nucleic Acid ◽  
Hcv Rna ◽  
Chimeric Mice ◽  
Rt Pcr ◽  
Modified Oligonucleotide

Background.In situhybridization (ISH) with high sensitivity has been requested to demonstrate hepatitis C virus (HCV) RNA in formalin-fixed, paraffin-embedded (FFPE) sections of the liver.Methods. ISH employing a locked-nucleic-acid- (LNA-)modified oligonucleotide probe and biotin-free catalyzed signal amplification system (CSAII) was applied to HCV-RNA detection in the liver tissue. Nested reverse-transcription polymerase chain reaction (RT-PCR) was performed for HCV genotyping using total RNA extracted from FFPE sections. The target tissues included FFPE tissue sections of humanized livers in HCV-infected chimeric mice (HCV genotypes 1a, 1b, and 2a and noninfected) and of needle-biopsied livers from HCV-infected patients.Results. HCV-RNA was demonstrated with the ISH technique in HCV-infected liver tissues from both chimeric mice and 9 (82%) of 11 patients with HCV infection. The HCV signals were sensitive to RNase. Nested RT-PCR confirmed the genotype in 8 (73%) of 11 livers (type 1b: 6 lesions and type 2a: 2 lesions). HCV-RNA was not identified in chronic hepatitis B lesions, fatty liver, autoimmune hepatitis, and hepatocellular carcinoma.Conclusion. ISH using the LNA-modified oligonucleotide probe and CSAII was applicable to detecting HCV-RNA in routinely prepared FFPE liver specimens.

scholarly journals Evidence of hepatitis in patients receiving transfusions of blood components containing antibody to hepatitis C

Blood ◽  
1993 ◽  
Vol 82 (3) ◽  
pp. 1000-1005 ◽  
Author(s):  
SK Aoki ◽  
PV Holland ◽  
LP Fernando ◽  
IK Kuramoto ◽  
S Anderson ◽  
...  
Keyword(s):  
Hepatitis C ◽  
Hcv Infection ◽  
Hcv Rna ◽  
Blood Components ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Immunoblot Assay ◽  
Polymerase Chain ◽  
Hepatitis C Virus Antibody

Abstract When hepatitis C virus antibody (anti-HCV) enzyme immunoassay (EIA1) testing became available in 1990, we tested samples from previously transfused blood units, traced the recipients of reactive units, and evaluated the recipients for HCV infection during the 12 months after transfusion. Ten of 42 recipients of EIA1-reactive blood were anti-HCV reactive on follow-up by EIA1 and 12 were reactive by a second- generation assay (EIA2). Reverse transcriptase-polymerase chain reaction (RT-PCR) detected HCV RNA in 5 seronegative recipients. In all, 17 of 42 recipients (40%) of EIA1-reactive blood had evidence of HCV infection. In comparison, 54 surgery patients, who received either no transfusion or autologous EIA1-nonreactive blood, remained EIA1 nonreactive and RT-PCR negative for 1 year; 1 patient (1.8%) became EIA2 reactive (P < or = .01). Of the recipients of anti-HVC reactive blood transfusions (reactive by both EIA1 and a supplemental 4-antigen strip immunoblot assay [RIBA2]), 14 (93%) of the recipients had evidence of HCV infection compared with only 3 of 27 recipients (11%) of EIA1-reactive, RIBA2-nonreactive blood (P < or = .01). Thus, blood components reactive for anti-HCV EIA1 may have transmitted HCV up to 40% of the time, but blood components found reactive by both EIA1 and RIBA2 may transmit HCV with a frequency of greater than 90%.

scholarly journals Long-term effects of hepatitis C virus infection in allogeneic bone marrow transplant recipients

Blood ◽  
1995 ◽  
Vol 86 (4) ◽  
pp. 1614-1618 ◽  
Author(s):  
P Ljungman ◽  
N Johansson ◽  
J Aschan ◽  
H Glaumann ◽  
B Lonnqvist ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Liver Dysfunction ◽  
Allogeneic Bone Marrow Transplantation ◽  
Allogeneic Bone Marrow ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hcv Rna ◽  
Allogeneic Bone

A total of 161 patients transplanted between 1978 and 1991 and who had survived at least 2 years after allogeneic bone marrow transplantation (BMT) were studied. Of 161 surviving patients, 28 (17.4%) were positive for hepatitis C virus (HCV) either by serology or polymerase chain reaction (PCR). Twenty-five patients were positive for HCV RNA by PCR, and 26 of the 28 patients had HCV antibodies detected by enzyme-linked immunosorbent assay (ELISA). The median follow-up time of HCV-positive patients was 6.1 years (range, 2.8 to 14.0 years). There was no difference in the frequency or degree of liver dysfunction between patients who were PCR-positive or -negative before BMT. Six patients developed severe liver dysfunction after BMT, and five of these patients did so after discontinuation or tapering of immunosuppression. No patient has developed liver failure. Serum transaminases were abnormal at the time of last follow up in 19 of 28 (68%) patients. Fifteen patients have had liver biopsies. No biopsy showed development of cirrhosis. We conclude that HCV is not a major contributing factor to morbidity and mortality during the first 5 to 10 years after allogeneic BMT.

scholarly journals Inhibitory Effect of a Triterpenoid Compound, with or without Alpha Interferon, on Hepatitis C Virus Infection

2011 ◽  
Vol 55 (6) ◽  
pp. 2537-2545 ◽  
Author(s):  
Takako Watanabe ◽  
Naoya Sakamoto ◽  
Mina Nakagawa ◽  
Sei Kakinuma ◽  
Yasuhiro Itsui ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Real Time ◽  
Interferon Regulatory Factor ◽  
Mechanisms Of Action ◽  
Alpha Interferon ◽  
Regulatory Factor ◽  
Rt Pcr ◽  
Response Element ◽  
Expression Levels

ABSTRACTA lack of patient response to alpha interferon (α-IFN) plus ribavirin (RBV) treatment is a major problem in eliminating hepatitis C virus (HCV). We screened chemical libraries for compounds that enhanced cellular responses to α-IFN and identified a triterpenoid, toosendanin (TSN). Here, we studied the effects and mechanisms of action of TSN on HCV replication and its effect on α-IFN signaling. We treated HCV genotype 1b replicon-expressing cells and HCV-J6/JFH-infected cells with TSN, with or without α-IFN, and the level of HCV replication was quantified. To study the effects of TSN on α-IFN signaling, we detected components of the interferon-stimulated gene factor 3 (ISGF3), phosphorylated signal transducer and activator of transcription 1 (STAT1), and STAT2 by Western blotting analysis; expression levels of mRNA of interferon regulatory factor 9 using real-time reverse transcription-PCR (RT-PCR); and interferon-stimulated response element reporter activity and measured the expression levels of interferon-inducible genes for 2′,5′-oligoadenylate synthetase, MxA, protein kinase R, and p56 using real-time RT-PCR. TSN alone specifically inhibited expression of the HCV replicon (50% effective concentration = 20.6 nM, 50% cytotoxic concentration > 3 μM, selectivity index > 146). Pretreatment with TSN prior to α-IFN treatment was more effective in suppressing HCV replication than treatment with either drug alone. Although TSN alone did not activate the α-IFN pathway, it significantly enhanced the α-IFN-induced increase of phosphorylated STATs, interferon-stimulated response element activation, and interferon-stimulated gene expression. TSN significantly increased baseline expression of interferon regulatory factor 9, a component of interferon-stimulated gene factor 3. Antiviral effects of treatment with α-IFN can be enhanced by pretreatment with TSN. Its mechanisms of action could potentially be important to identify novel molecular targets to treat HCV infection.

scholarly journals Performance Evaluation of Commercial Dengue Diagnostic Tests for Early Detection of Dengue in Clinical Samples

2017 ◽  
Vol 2017 ◽  
pp. 1-4 ◽  
Author(s):  
Tuan Nur Akmalina Mat Jusoh ◽  
Rafidah Hanim Shueb
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Diagnostic Test ◽  
Rapid Diagnostic Test ◽  
Dengue Infection ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Kappa Agreement

The shattering rise in dengue virus infections globally has created a need for an accurate and validated rapid diagnostic test for this virus. Rapid diagnostic test (RDT) and reverse transcription-polymerase chain reaction (RT-PCR) diagnostic detection are useful tools for diagnosis of early dengue infection. We prospectively evaluated the diagnostic performance of nonstructural 1 (NS1) RDT and real-time RT-PCR diagnostic kits in 86 patient serum samples. Thirty-six samples were positive for dengue NS1 antigen while the remaining 50 were negative when tested with enzyme-linked immunosorbent assay (ELISA). Commercially available RDTs for NS1 detection, RTK ProDetect™, and SD Bioline showed high sensitivity of 94% and 89%, respectively, compared with ELISA. GenoAmp® Trioplex Real-Time RT-PCR and RealStar® Dengue RT-PCR tests presented a comparable kappa agreement with 0.722. The result obtained from GenoAmp® Real-Time RT-PCR Dengue test showed that 14 samples harbored dengue virus type 1 (DENV-1), 8 samples harbored DENV-2, 2 samples harbored DENV-3, and 1 sample harbored DENV-4. 1 sample had a double infection with DENV-1 and DENV-2. The NS1 RDTs and real-time RT-PCR tests were found to be a useful diagnostic for early and rapid diagnosis of acute dengue and an excellent surveillance tool in our battle against dengue.

Performance of two Real-Time RT-PCR assays for quantitation of hepatitis C virus RNA: Evaluation on HCV genotypes 1-4

2010 ◽  
Vol 82 (11) ◽  
pp. 1878-1888 ◽  
Author(s):  
Abeer Elkady ◽  
Yasuhito Tanaka ◽  
Fuat Kurbanov ◽  
Fuminaka Sugauchi ◽  
Masaya Sugiyama ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Real Time ◽  
Rt Pcr ◽  
Hepatitis C Virus Rna ◽  
Hcv Genotypes ◽  
Virus Rna ◽  
Pcr Assays

scholarly journals Prevalence of Bovine viral diarrhea virus in cattle herds from Basrah and Nassirya Provinces by direct and indirect Elisa and Real time qPCR

2012 ◽  
Vol 11 (2) ◽  
pp. 1
Author(s):  
B. A. Jarullah, J. Aed Gati, and A. Saleh
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Indirect Elisa ◽  
Positive Sample ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Diarrhea Virus ◽  
Virus Rna ◽  
Two Samples ◽  
Cattle Herds

The current study was conducted to investigate the prevalence of BVD virus in Basrah and Nassirya city by using ELISA and RT-PCR. Two hundreds and eighty two samples of non vaccinated cattle sera samples collected from two regions of Iraq (188 samples from Nassirya city and 92 samples from Basrah city). Samples tested by Enzyme Linked Immunosorbent Assay (ELISA) antigen capture. Positive results were 20 samples ( 8 sample in Thi-Qar and 12 positive samples from Basrah). All samples submitted to indirect ELISA(IDEXX HerdCheck ELISA )for detect BVDV antibodies .Genotyping of all 20 positive samples to antigen detection were tested by Real time PCR, using Cador BVDV ½ kit, after extraction of virus RNA by QIAamp mini kit. The results revealed that there were 20 positive sample according to direct ELISA(Ag detection), while 66 sample were positive to indirect ELISA, as well as, the result of RT-PCR showed that there were two sample positive to BVDV type-1 (one sample form each city).Key words: BVDV, Genotype, ELISA, Iraq, Real time PCR.

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