detection linearity and reproducibility of hbv cccdna real time pcr

... performance of the PCR was monitored by quantitative real- time PCR (qPCR) The mean cycle threshold (Ct) and the standard deviation of the controls were calculated Samples in which the mean Ct of the test ... are conducted in a single PCR and therefore the chances of possible contamination are minimised The main advantage of real- time detection is the large dynamic range offered in a quantitative ... cycles Standards of 106 to 10 copies per reaction of C trachomatis pgp3 amplicons and negative controls were included in each PCR reaction to generate a standard curve and quantities of the unknown...

Ngày tải lên: 19/06/2014, 08:20

... performance of the PCR was monitored by quantitative real- time PCR (qPCR) The mean cycle threshold (Ct) and the standard deviation of the controls were calculated Samples in which the mean Ct of the test ... are conducted in a single PCR and therefore the chances of possible contamination are minimised The main advantage of real- time detection is the large dynamic range offered in a quantitative ... cycles Standards of 106 to 10 copies per reaction of C trachomatis pgp3 amplicons and negative controls were included in each PCR reaction to generate a standard curve and quantities of the unknown...

Ngày tải lên: 20/06/2014, 04:20

... et al Detection limit and standard curve of the multiplex real- time PCR The detection limit and standard curve of the multiplex real- time PCR was determined using S Typhimurium ATCC 14028 and S ... the results of the standard culture method of isolated field and reference strains with those of the multiplex real- time PCR Results Specificity of the multiplex real- time PCR A total of 128 bacterial ... previous real- time PCR assays [8,27] These advantages improved the multiplex real- time PCR assay in terms of shortening turnaround time for bacterial detection and reducing the risk of crosscontamination...

Ngày tải lên: 07/08/2014, 23:22

... use Page of and PCV-2), and control (sterile water) were processed with the real- time PCR Reproducibility of real- time PCR To determine the reproducibility of the real- time PCR, the standard plasmid ... sensitivity of the real- time PCR is 100 times greater than that of the conventional PCR (Figure 1C) Specificity of the real- time PCR The real- time PCR gave positive results for the standard plasmid of ... and 72°C for 20 s The data were analyzed with the Rotor-Gene software Sensitivity of the real- time PCR To determine the detection limit and efficiency of the assay, recombinant plasmid of standard...

Ngày tải lên: 12/08/2014, 02:20

... Green Real Time PCR TaqMan Real time PCR reaction Real time PCR reactions that target the E1 region of HPV were performed using Roche Lightcycler Sybr Green master mix Each reaction consisted of ... E1 region of known HPVs and should allow for detection of any of these types The second qPCR assay targets the coding region of the hypervariable loop V of the L1 gene of HPV6, 11, 16 and 18 These ... quantitation of viral DNA to address the role of viral load in disease progression Real time quantitative PCR (qPCR) allows for quantitation of DNA over orders of magnitude [11] While qPCR assays...

Ngày tải lên: 12/08/2014, 04:20

... efficient one-step RT -PCR assay for CHIKV detection and quantification The additional advantage of utilizing SYBR Greenbased real- time RT -PCR is that it is relatively easy to design and test primer ... that of the one-step SYBR Green-based RT -PCR assay The obvious amplified 107bp DNA bands can only be observed to the detection limit of 103 PFU/ml of CHIKV Immunofluorescence assay (IFA) detection ... sensitivity of the assay in the detection of CHIKV infection To verify the specificity of the nsP2 primers in the current one-step SYBR Green-based real- time RT -PCR, amplification of RNA extracted...

Ngày tải lên: 12/08/2014, 04:21

... resulted in the presence of Ct value in real- time PCR detection The detection limit of the conventional PCR was determined through the highest dilution that resulted in the presence of clear amplified ... TaqMan real- time PCR for detection of hepatopancreatic parvovirus from Australia J Virol Methods 2007, 140:10-16 42 Hussein IT, Field HJ: Development of a quantitative real- time TaqMan PCR assay ... and application of a one-step real- time Taqman RT -PCR assay for detection of Duck hepatitis virus type1 J Virol Methods 2008, 153:55-60 Deng S, Cheng A, Wang M, Cao P: Serovar-specific real- time...

Ngày tải lên: 12/08/2014, 04:21

... primer sets for detection of all four serotypes of DENV [11] In the present study, we describe the development of a DENVspecific TaqMan based real- time PCR for detection and quantitation of all four ... directly for sequencing and for evaluation of sensitivity of the real- time PCR JE, WN and CHK viruses were used to evaluate the specificity of the assay Infection and maintenance of cell cultures Vero ... dengue Another important aspect of dengue disease is the surveillance of vector population and detection of DENV in field caught mosquitoes Real- time PCR because of its high sensitivity could be...

Ngày tải lên: 12/08/2014, 04:21

... Features of ATP III similar to those of ATP I and II Identification of population groups, besides middle-aged men, for detection of high LDL cholesterol (and other lipid risk factors) and for ... Ref–1 xii Detection Evaluation Treatment I Background and Introduction I Background and Introduction The Third Report of the Expert Panel on Detection, Evaluation, and Treatment of High Blood ... as target of therapy Notwithstanding this diverse evidence, the ultimate proof of the benefits of lowering LDL cholesterol is through clinical trial A large number of clinical trials of cholesterol-lowering...

Ngày tải lên: 14/02/2014, 09:20

... VZV strain Schenke and HIV-1 strains BaL, NL4.3 and RFW were propagated according to standard protocols [16-20] The cell types infected, MOI, time of infection and percentage of cells infected ... harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used QPCR normalisation genes in both and mock and virally infected cells ... gene for use in QPCR studies of viral infections Nevertheless it is of benefit for researchers to be able to determine a shortlist of potential candidates To date the variability of housekeeping...

Ngày tải lên: 18/06/2014, 18:20

... expressed as copies of viral DNA per 100 µL of virus suspension The reliability of real- time PCR was defined by calculating coefficients of variation of Ct values of replicates of standard curve dilutions ... Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral load A – C: Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral ... microtiter and transwell plates Semiquantitative polymerase chain reaction (PCR) and quantitative real- time PCR have been recently developed and employed for the detection and quantitation of JCV...

Ngày tải lên: 19/06/2014, 08:20

... VZV strain Schenke and HIV-1 strains BaL, NL4.3 and RFW were propagated according to standard protocols [16-20] The cell types infected, MOI, time of infection and percentage of cells infected ... harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used QPCR normalisation genes in both and mock and virally infected cells ... gene for use in QPCR studies of viral infections Nevertheless it is of benefit for researchers to be able to determine a shortlist of potential candidates To date the variability of housekeeping...

Ngày tải lên: 20/06/2014, 01:20

... expressed as copies of viral DNA per 100 µL of virus suspension The reliability of real- time PCR was defined by calculating coefficients of variation of Ct values of replicates of standard curve dilutions ... Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral load A – C: Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral ... microtiter and transwell plates Semiquantitative polymerase chain reaction (PCR) and quantitative real- time PCR have been recently developed and employed for the detection and quantitation of JCV...

Ngày tải lên: 20/06/2014, 04:20

... treatment of ovarian cancer has come from intraperitoneal administration of platinum and taxane agents instead of the more conventional intravenous delivery of these drugs [71-73] Of the 654 randomized ... 2Department of Obstetrics and Gynecology, University of Wisconsin-Madison, 600 Highland Ave, Madison, WI, 53792, USA Authors’ contributions JAAG, JPC, and MSP did the majority of the writing of this ... I-chainrelated proteins A and B (MICA/MICB) and the UL16binding proteins (ULBP-3) [105] NKG2D ligands are not expressed on normal, healthy cells and therefore the expression of NKG2D ligands is correlated...

Ngày tải lên: 20/06/2014, 07:20

... Face detection and tracking Evaluation of the face detection and tracking is accomplished using an extensive array of experimental data We collected many video clips of different setups and in ... with DFFS bounds of 2500 and 4000 We have used our real- time videos for face detection and tracking evaluations Note that it is also possible to test the single-frame face searching and verification ... on Image and Video Processing (a) (b) (c) Figure 10: Samples of indoor and outdoor test video clips for counting the face detection rates and false positives Table 1: Face detection and false...

Ngày tải lên: 22/06/2014, 19:20

... for the measurement of the average molecular weight of HMW hyaluronan Analysis of message expression by quantitative realtime RT -PCR Message expression in the synovium of knees and the relative ... levels of the messages (RA) Relative expressionand rheumatoid arthritis for hyaluronanof knees in 2, and -3 and hyaluronidase-1, -2, and -3 in the synovium of knees in osteoarthritis (OA) and rheumatoid ... the messages of both target gene and β-actin gene in the same sample tube when the intensity of the reporter fluorescent signal reached 10 times the standard deviation of the baseline of fluorescent...

Ngày tải lên: 09/08/2014, 01:24

... and extraction of genetic material Real time- quantitative polymerase chain reaction The levels of E-cadherin, α-, β- and γ-catenin transcripts from the prepared cDNA were determined by real time ... different cut-offs for positivity and negativity We determined the expression of E-cadherin, α-, β- and γcatenin in human breast carcinomas using real time quantitative polymerase chain reaction and investigated ... that allows real- time detection of 96 reactions was used to amplify the plasmid standards and breast tissue samples under the following conditions: 94°C for 12 minutes; 50 cycles of 94°C for...

Ngày tải lên: 09/08/2014, 07:21

... detecting and defining homozygous segments consists of: 1) basic detection of runs of homozygous and heterozygous genotypes; 2) joining of neighbouring homozygous segments across regions of low ... Comparison of the extent and frequency of homozygous segments with haplotypes underlying extAUC peaks Analysis of the consistency of the homozygous extent distribution and length and frequency of haplotypes ... samples and CEU (r = 0.73 and 0.74 with CHB and JPT, respectively), and low-moderate correlation between YRI and the other three population samples (r = 0.64, 0.53, and 0.56 for CEU, CHB, and JPT,...

Ngày tải lên: 09/08/2014, 22:23

... control and PCV1, PRRS, PED, TGE and RV samples Detection of clinical samples Table and showed that the PCV2-positive rates in the unknown samples of conventional PCR detection and real- time PCR detection ... provide an alternative approach for detection of PCV2 Table Comparison between conventional PCR and real- time PCR for PCV2 positive samples Conventional PCR Real time PCR Subtotal + - + 3 - 0 Total ... correlation (R2) of 0.9999 between the Ct value and the logarithm of the plasmid copy number LOD and LOQ of the assay TaqMan real- time PCR Real- time PCR was carried out on an ABI 7500 thermocycler...

Ngày tải lên: 11/08/2014, 21:21

... by a dissociation curve and agarose gel electrophoresis Real- time PCR conditions and analysis PCR reactions were performed in 96-well plates with the Chromo4 Real- Time PCR Detector System (BioRadđ) ... apomictic accession, with 98% of aposporous embryo sacs [9] Quantitative real- time PCR (qRT -PCR) has emerged as an important technique to compare the expression profiles of target genes in different ... treatments and also to validate high-throughput gene expression profiles [15,16] One of the methodologies to determine gene expression levels in qRT -PCR is by comparing the expression of the gene of...

Ngày tải lên: 12/08/2014, 03:20