detection of hbv cccdna methylation from clinical samples by bisulfite sequencing and methylation specific pcr
Methods in molecular biology vol 1540 hepatitis b virus methods and protocols
... Daniela Stadler, Chunkyu Ko, and Ulrike Protzer Detection of HBV cccDNA Methylation from Clinical Samples by Bisulfite Sequencing and Methylation- Specific PCR Yongmei Zhang, Richeng ... effects on intracellular calcium metabolism by the Bouchard lab; detection, cloning, and sequencing of HBV markers in laboratory-generated and clinical samples by Dandri, Huang, Jilbert, Weiland, and ... DNA sequence 3.8 Examining HBV cccDNA Wash HepG2-NTCP cells in a 48-well plate with 200 μL PBS once on day postinfection 3.8.1 Quantification of HBV cccDNA by qRT -PCR Add 200 μL cell lysis buffer...
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Báo cáo y học: "Epidemiology and Prevention of Hepatitis B Virus Infection"
... all countries with a hepatitis B carrier prevalence (HBsAg) of 8% or greater by 1995 and in all countries by 1997 By May 2002, 154 countries had routine infant immunization with hepatitis B vaccine ... progress HBeAg-negative CHB HBeAg-negative chronic hepatitis B (e-CHB), characterized by HBV DNA levels detectable by nonamplified assays and continued necroinflammation in the liver, has been reported ... [20,22]; (2) cellular transfer: the placental tissue is infected by hightiter of HBV in maternal blood from mother’s side to fetus’ step by step, and finally, HBV reach fetus’ circulation through the...
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Báo cáo y học: "Natural History and Clinical Consequences of Hepatitis B Virus Infection"
... with precore HBV [8] During the treatment of chronic hepatitis B by lamivudine, drug resistance may develop, which is mediated by point mutations with the YMDD motif at the catalytic center of ... 10 weeks of incubation period, followed by antibodies against the core antigen (HBcAb) in IgM form during early period [2] Viremia is well established by the time HBsAg is detected (usually from ... suggested positive HBsAg increased one’s risk of developing HCC by 10 folds, and with positive HBeAg, HCC is significantly increased by 60 folds Moreover, a detectable HBV DNA level yields a fold...
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Báo cáo khoa học: "Use of dried blood samples for monitoring hepatitis B virus infection" ppsx
... b) Amplification by PCR In order to determine the utility of the extracted nucleic acids from DBS samples for molecular studies, the DNA extracted was utilized as a template for PCR to amplify ... temperatures and plasma samples for HBV -PCR amplification was optimal, because all the samples were successfully amplified (Fig 1) c Correlation between HBV-DNA VL in DBS samples stored at different ... significant by the ANOVA test The CI 95% for the gold standard in plasma was (3.1-7.83), for DBS at: -20 Figure of Stability the dried blood samples Stability of the dried blood samples PCR amplifications...
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siRNA, miRNA and their possible roles in molecular therapy of chronic human hepatitis b virus infection and hepatocellular carcinoma
... Table 3.2 Sequences of PCR primers and probes for real-time PCR quantification of HBV DNA …………………………………………………… 64 Table 3.3 Sequences of PCR primers and probes for real-time PCR quantification of ... Livers ……… ……… ……… ……… 115 116 5.2.1 Choice of Patient Samples and Validation of RT-real-time PCR Approach… 117 5.2.1.1 Selection of patient RNA samples …………………….….…… 117 5.2.1.2 Quality of RNA ... of HBV Gene Expression by siRNAs 4.1 Introduction…………………………………………………………….… …… 81 4.2 Selection and Transfection of HBV Specific siRNAs………………… 82 4.2.1 Selection of HBV Specific siRNAs………………………...
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Báo cáo y học: "Enhanced surveillance for childhood hepatitis B virus infection in Canada, 1999-2003"
... goodness-of-fit statistics on the residual deviance Differences in proportions were determined by a chi-square test and, where necessary, Fisher’s exact test The Kruskal-Wallis test was used ... acquiring HBV infection (RR 3.56, 95% CI 1.57-8.09) DISCUSSION Within those health regions covered by the EHSSS, the Public Health Agency of Canada supports staff in conducting enhanced surveillance, ... notification system was not materially changed during the study period, but augmented throughout by scrutiny of laboratory results and hospital admissions; therefore the incidence trend was not...
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Báo cáo khoa học: " Selective receptor expression restricts Nipah virus infection of endothelial cells" pot
... visualized by DAPI staining Magnification, ×100 (B) mRNA was extracted from × 105 cells by standard procedures and subjected to RT -PCR with EB2 -specific primers (C) RT -PCR with EB3 -specific primers ... assumed to be expressed on EC in vivo, expression of EB3 in our model EC was analyzed by an EB3 -specific RT -PCR [33] Fig 2C shows that besides Vero cells, only PBMEC express small amounts of EB3 ... with NiV (MOI of 0.2), visualized virus-positive cells by immunostaining at 24 h p.i and analyzed virus release into the supernatant by RT -PCR at 48 h p.i In contrast to wildtype PAEC, PAEC-EB2...
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Pharmacoeconomics and health outcomes research for hepatitis b virus infection in singapore
... general population On the other hand, disease -specific HRQoL instruments comprise items that aim to measure areas of life specifically affected by the disease of interest (Fayers and Machin, ... sensitivity analysis by varying the cost of management and fix the estimated number of patients in different disease stages……………………………………………………………… 122 Table 5.3 One- way sensitivity analysis by varying ... analysis by varying the indirect cost (25th 75th percentile) for each HBV disease state and fix the estimated number of patients……………………………………………… 133 Table 5.11 One-way sensitivity analysis by varying...
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báo cáo hóa học:" High Prevalence of Hepatitis B Virus Markers in Romanian Adolescents With Human Immunodeficiency Virus Infection" ppt
... lower limit of detection of 1000 HBV DNA copies/mL HIV Infection Markers Virologic and immunologic HIV infection markers were determined, each by an independent investigator, only for samples collected ... surveillance after a rubella catchup vaccination program Whole blood was collected by venipuncture into EDTA anticoagulant Samples were centrifuged within hours after collection, and plasma was transferred ... HIV RNA copies/mL (lower detection limit, 400 copies/mL; linear range, 400750,000 copies/mL) Statistical Analysis Differences in HBV marker prevalence rates were evaluated by Fisher's exact test...
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Báo cáo y học: " Prevalence of hepatitis delta virus infection among hepatitis b virus surface antigen positive patients circulating in the largest province of pakistan" pptx
... these, 80 anti-HDV ELISA positive samples were further analyzed by HDV PCR Total 56 anti-HDV ELISA positive samples were found negative for HDV RNA by PCR All the samples belonged to different regions ... used in nested PCR Qualitative detection of HDV cDNA The qualitative detection of HDV-cDNA was carried out by nested PCR using four μl of synthesized HDV-cDNA The first round PCR was done in ... ELISA positive samples, total 80 (83%) samples were found reactive by anti-HDV ELISA method Of these HBsAg and anti-HDV positive samples, 24 (30%) were further found positive by HDV RNA PCR Zaidi...
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Báo cáo y học: " Seropositivity of Hepatitis B virus and Hepatitis C virus dual Infection among blood donors in Nyala Teaching Hospital" pot
... Microbiology 2004, 22(2):136-137 Pallas JR, Farinas-Alvarez C, Prieto D, Delgado-Rodriguez M: Coinfection by HIV, Hepatitis B and Hepatitis C in imprisoned injecting drug users Europe Journal of Epidemiology...
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Báo cáo y học: "Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) Dual Infection"
... both HBsAg and antiHBs might be a group of patients with a multiple HBV and HCV infection HBV DNA by PCR was detected in 40.8% of 71 such patients in this study, which implies that nearly half of ... enhancer up to 11-fold, the enhancer 3-4-fold Suppression of HBV enhancer by HCV core from genotype 1b was stronger than by HCV core of genotypes 3a or 1a This trans-repression may contribute to ... 30.4%, p=0.009) Similar results have been observed by others in patients having silent HBV coinfection with HCV [48] It has been suggested that a specific dose and duration of IFN regimen for the...
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Báo cáo y học: "Study of the early steps of the Hepatitis B Virus life cycle"
... confirmed by reverse-transcription PCR (RT -PCR) and Northern blot analysis As shown in figure 2, two specific primer sets, PI1/ PI2 and PI3/PI4, were used to amplify SPIK mRNA by RT -PCR PI1/PI2 ... 1(1): 21-33 32 Figure RT -PCR detects SPIK RNA 1µg RNA from HepG2 cells and human liver cells were reversely transcripted and amplified by PCR using specific primer for SPIK PCR bands were resolved ... membrane, and then hybridized with a 32P labeled SPIK specific probe derived from the SPIK gene by PCR In agreement with the results from RT -PCR, figure shows that the SPIK mRNA can only be detected...
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Báo cáo khoa học: Betulinic acid-mediated inhibitory effect on hepatitis B virus by suppression of manganese superoxide dismutase expression pot
... * * (Arbitrary units) 100 pCREB level by Western blot (Arbitrary units) 200 pCREB (Ser133) level by Western blot BetA/CREB 200 siCREB C BetA 300 SOD2 protein level by Western blot (Arbitrary ... (Arbitrary units) B BetA * * CTL CREB protein level by Western blot * SOD2 mRNA level by qPCR (Arbitrary units) 150 (Arbitrary units) qPCR on SOD2 promoter by CREB ChIP A * 100 *¶ CTL BetA BetA/CREB siCREB ... subsequent ROS generation E 200 HBV DNA level by qPCR (Arbitrary units) B SOD2 mRNA level by Western blot (Arbitrary units) A SOD2 mRNA level by qPCR (Arbitrary units) cells; as shown in Fig 5E,...
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Báo cáo khoa học: Expression studies of the core+1 protein of the hepatitis C virus 1a in mammalian cells pot
... mediated by slippage of ribosomes during translation elongation at core codons 8–11 and yields a core+1 chimeric protein containing the first 8–11 amino acids of core fused to amino acids encoded by ... core+1 protein remains largely unknown, as functional studies of the core+1 ORF are limited by the elusive detection of its native form in cultured cells expressing the HCV structural region or in ... that the form of the core+1 protein produced by frameshift (core+1 ⁄ F), is a short-lived protein whose half-life could be substantially increased by the addition of chemical proteasome inhibitors...
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Báo cáo khoa học: Fidelity of hepatitis B virus polymerase pptx
... reaction was terminated by the addition of EDTA to a final concentration of 50 mM in 95% formamide buffer Reaction products were denatured by incubating at 95 °C for and analyzed by electrophoresis ... microliters of 95% formamide were added and the proteins were immediately denatured by incubating at 95 °C for and analyzed by electrophoresis in M urea/16% polyacrylamide gels The gel was then dried ... using T4 polynucleotide kinase (20 U) Reaction was started by adding 100 nM oligonucleotide and incubated at 37 °C for 1.5 h, and stopped by adding EDTA to the final concentration of 20 mM The reaction...
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Báo cáo sinh học: " Common Genotypes of Hepatitis B virus prevalent in Injecting drug abusers (addicts) of North West Frontier Province of Pakistan" doc
... Suppl):S158-S168 Naito H, Hayashi S, Abe K: Rapid and specific genotyping system for hepatitis B virus corresponding to six major genotypes by PCR using type specific primers Journal of Clinical Microbiology ... IL, USA Molecular analysis Serological positive samples for HBsAg were further processed for HBV genotypes by the same methodology using genotype specific primers as described previously28 Briefly, ... virus M: 50 bp marker; Lane 2–3: HBV genotype A specific 68 bp band; Lane 4–5: HBV genotype D specific 119 bp band; Lane 6–10: genotype A and D specific 68 bp and 119 bp bands in patients with...
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Báo cáo sinh học: " Hepatitis B virus (HBV) genotypes in Egyptian pediatric cancer patients with acute and chronic active HBV infection" pptx
... were determined by sequencing and found that HBV genotype D are most prevalent in Egypt [14] Naito et al (2001) examined serum samples positive for HBV DNA by primer specific PCR and these turned ... (type D specific) , BE1 (type E specific) , and BF1 (type F specific) These primer combinations for second-round PCR were designed on the basis of the differences in the sizes of the genotype -specific ... serum samples from Egyptian carriers were determined by sequencing and found that HBV genotype D are most prevalent in Egypt [14] Naito et al (2001) examined serum samples positive for HBV DNA by...
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Báo cáo sinh học: " Serum levels of preS antigen (HBpreSAg) in chronic hepatitis B virus infected patients" potx
... Quantification of HBV DNA in serum by PCR Quantification of serum HBV DNA was performed by the LightCycler PCR system (Roche, Basel, Switzerland) using fluorescent quantitative PCR kits (PG Biotech, Shenzhen, ... specimens, with reconciliation by HBeAg results HBV+ (DNA+ reconciled by HBeAg+) HBV- (DNA- reconciled by HBeAg-) 52 12 30 55 HBpreSAg+ HBpreSAg- Sensitivity(%) CI,95% Specificity(%) PPV(%) NPV (%) ... subtracting the background below 20 µg/ml, samples were diluted by 2X and 10X for HBeAg-negative patients and HBeAg-positive cases, respectively All samples were tested in duplicate and the average...
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Báo cáo sinh học: " Cellular apoptosis induced by replication of hepatitis B virus: possible link between viral genotype and clinical outcome" pptx
... 1:5000 dilution Following ECL detection (Pierce, USA) membrane was developed by Kodak E&D system (Kodak, USA) Alignment analysis HBSP amino acid sequences were analyzed by using Vector NTI9 Result ... apoptosis supported by FACS analysis Further investigation identified the natural expression of HBSP in HBV genome transfected cells The observed difference in apoptotic effect caused by the three HBV ... vector and cells treated by 50 µM cisplatin for 16 hr were set as (-) and (+) controls Transfected cells were collected at 24 hr and 48 hr after incubation and analyzed by Apoalert™ annexin-V kit...
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