lod of the two duplex real time pcr assays

... real- time PCR standard curve Establishment of the fluorescent quantitative real- time PCR standard curve Standard curve of the AHV-1 fluorescent quantitative real- time PCR Ten-fold dilutions of standard ... fluorescent quantitative real- time PCR and conventional PCR The real- time PCR was carried out using the ABI AmpliTaq Gold DNA polymerase system with an icycler IQ Realtime PCR Detection System (Bio-Rad ... improvement of the previously developed ordinary TaqMan real- time PCR method Results Development and optimization of FQ -PCR and conventional PCR Following the optimization of FQ -PCR, final concentrations...

Ngày tải lên: 12/08/2014, 04:21

... real- time PCR standard curve Establishment of the fluorescent quantitative real- time PCR standard curve Standard curve of the AHV-1 fluorescent quantitative real- time PCR Ten-fold dilutions of standard ... fluorescent quantitative real- time PCR and conventional PCR The real- time PCR was carried out using the ABI AmpliTaq Gold DNA polymerase system with an icycler IQ Realtime PCR Detection System (Bio-Rad ... improvement of the previously developed ordinary TaqMan real- time PCR method Results Development and optimization of FQ -PCR and conventional PCR Following the optimization of FQ -PCR, final concentrations...

Ngày tải lên: 12/08/2014, 04:21

... optimization of the Duplex Real- time PCR condition and evaluation of the performance of this method through determining the sensitivity and specifictity The results indicated that the DNA extraction ... ảnh hưởng đến phản ứng Duplex Real- time PCR 30 Bảng 3.3 Thành phần phản ứng Single Real- time PCR 30 Bảng 3.4 Chương trình nhiệt phản ứng Single Duplex Real- time PCR 31 Bảng 3.5 Thành ... 3.5 Thành phần phản ứng Single Real- time PCR 31 Bảng 3.6 Thành phần phản ứng Duplex Real- time PCR 32 Bảng 3.7 Thành phần phản ứng Duplex Real- time PCR tối ưu 33 Bảng 3.8 Phân...

Ngày tải lên: 26/05/2018, 13:34

... instructions designed for the study The viral results were merged with HBV serological results prior to data analysis Results Performance of the real- time qPCR The performance of the new qPCR assay was examined ... Figure Comparison of real time PCR with the Roche Amplicor Monitor in 10 HBsAg positive samples The log difference between the two assays was less than 1.0 over the dynamic range of 1.1 × 103 to ... adoption of the methodology [15] More recently the development of real time PCR methodology has further improved the ease with which HBV DNA levels can be monitored and has increased the range...

Ngày tải lên: 20/06/2014, 01:20

... copies/μL The sensitivity of the real- time PCR was compared with conventional PCR (Yue et al., 2009) Specificity of the real- time PCR To determine the specificity of the real- time PCR, the standard ... 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 PPV strains ... Figure 1B The amplification product was about 123 bp long, and no false amplification was observed Sensitivity of the real- time PCR The detection limit of the real- time PCR for the NS1 gene of PPV...

Ngày tải lên: 12/08/2014, 02:20

... geometric mean of the expression values of the set of the control genes tested The lower the M value, the more stably expressed the gene is Also, the program enables the exclusion of the most unstable ... region of the gene The M value for both sets of primers was the same, showing that the amplification region had no influence on the expression stability Although the difference in the M value of the ... M value of all the other genes and also a change in the M values of the unstable genes, BbrizGDP and BbrizUBI (Figure 3b) To check if the Figure Box-whisker showing the Ct variation of each candidate...

Ngày tải lên: 12/08/2014, 03:20

... international standard units (IU/mL) Comparison of the ultra sensitive RTQ -PCR vs the previous real- time PCR assay To compare the ultra sensitive RTQ -PCR assay with the previously reported test, HBV-DNA ... values of the estimated (copies/mL) versus the expected values (IU/mL) and also between HBVDNA quantifications using the two in house real- time PCR assays and COBAS TaqMan as well To compare the ... sensitive RTQ -PCR and the previous test was: log10 HBV-DNA (copies/mL) with the ultra sensitive RTQ -PCR = -0.531 + 1.015x log 10 (copies/mL) with the previous real- time PCR test The 95% CI for the estimated...

Ngày tải lên: 12/08/2014, 04:20

... correct The LOD of the duplex real- time RT -PCR assay was 38.99 IU/ml and the specificity was 100% Furthermore, the cost of the duplex real- time RT -PCR assay was considerably lower than that of the ... H2O and used as the template for the duplex real- time RT -PCR assay Duplex real- time RT -PCR amplification for HCV RNA detection The duplex real- time RT -PCR assay was performed on the ABI PRISM system ... realtime RT -PCR assay Intrinsic performance of the duplex real- time RT -PCR assay Linearity Linearity of the duplex real- time RT -PCR assay was determined using serial 10-fold dilutions of a clinical...

Ngày tải lên: 12/08/2014, 04:20

... cells The real- time PCR (RT -PCR) technology utilizes the standard PCR method with the addition of measuring the accumulation of amplified DNA in real- time by a fluorescent signal RT -PCR uses the ... of the labeled, purified virus and C) MOI 0.05 TCID50/cell of unpurified virus incubated as described in A) and the amount of virus bound to the cell surface was measured with a two- step RT -PCR ... RD cells despite the absence of CAR [40] Using RT -PCR for sensitive studies of viral host cell interactions To explore the implementations of RT -PCR in viral binding studies the limitations in...

Ngày tải lên: 12/08/2014, 04:21

... Moreover, the microcystin-degrading bacteria in the biofilm of the biological treatment facility were quantified The bacteria were present in the biofilm throughout the year, suggesting that the biofilm ... real- time PCR It was found out that microcystin-degrading bacteria were present in the biofilm of the practical biological treatment facility throughout the year, and the abundance ratio of the ... in the biofilm in autumn at the practical biological treatment facility, but there is little information about the dynamics of the bacteria in lake water and other water bodies Therefore, further...

Ngày tải lên: 05/09/2013, 10:15

... networks and gene pathways To further understand the biological networks of the 19 genes, we next analyzed their biological interactions using the Ingenuity Pathways Analysis (IPA) tool Ten networks ... the principal component for the cell lines (PC1, PC2, and PC3), two of the three AD cells lines were close, whereas the third cell line was close in PC2 to the other two cell lines, but separate ... deposited in the CIBEX microarray database (accession CBX 100) Quantification of 19 candidate genes by qPCR Using qPCR, we quantified the expression of the 19 candidate genes in the same eight...

Ngày tải lên: 15/02/2014, 04:20

... genes, as any variation in the expression of the reference gene between sample groups will reduce the sensitivity of the assay to detect changes in the expression of genes of interest and may also ... normalisers for QPCR assays According to this analysis the most reliable overall reference gene was PP1A with a sumv value of 7.41 and the most reliable gene in of the 10 viral infections Other genes ... experiments and the reliability of a panel of potential genes should be determined for individual experimental conditions [3,8] Ideally the best two or three of these genes would then be used for...

Ngày tải lên: 18/06/2014, 18:20

... DNA The 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 ... in the CVL fluid of these 27 HSV PCR positive subjects The distribution is left censored and the majority of subjects had

Ngày tải lên: 19/06/2014, 08:20

... 60°C at the end of the cycle Real- time PCR amplification data were analyzed using the Bio-Rad iCycler iQ™ Multicolor Real- Time PCR Optical System Software Version 3.1 A standard curve for the quantitation ... 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 load JCV ... calculated from the standard curve and were 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...

Ngày tải lên: 19/06/2014, 08:20

... genes, as any variation in the expression of the reference gene between sample groups will reduce the sensitivity of the assay to detect changes in the expression of genes of interest and may also ... normalisers for QPCR assays According to this analysis the most reliable overall reference gene was PP1A with a sumv value of 7.41 and the most reliable gene in of the 10 viral infections Other genes ... experiments and the reliability of a panel of potential genes should be determined for individual experimental conditions [3,8] Ideally the best two or three of these genes would then be used for...

Ngày tải lên: 20/06/2014, 01:20

... DNA The 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 ... in the CVL fluid of these 27 HSV PCR positive subjects The distribution is left censored and the majority of subjects had

Ngày tải lên: 20/06/2014, 04:20

... 60°C at the end of the cycle Real- time PCR amplification data were analyzed using the Bio-Rad iCycler iQ™ Multicolor Real- Time PCR Optical System Software Version 3.1 A standard curve for the quantitation ... 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 load JCV ... calculated from the standard curve and were 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...

Ngày tải lên: 20/06/2014, 04:20

... compare 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 ... respectively For the specificity of the multiplex real- time PCR, the cut-off value was determined at a mean CT value of 30 Standard curve using pure cultures The multiplex real- time PCR assay was ... against the corresponding bacterial cell number (log10 CFU/ml) 47 Fig Standard curves of the multiplex real- time PCR for Salmonella (S.) Enteritidis The results of the multiplex real- time PCR were...

Ngày tải lên: 07/08/2014, 23:22

... periods of time Furthermore, the complexity of the studies limits the number of agents that can be tested Because of these issues, we have focused on the development of reliable biomarker assays ... content not shown) The horizontal axis in Fig shows the number of cells, while the vertical axis shows the C(t) where the PCR product could be detected When the efficiency of the Q -PCR reaction using ... amount of RNA Variability of direct C(t) determination versus the standard curve method To evaluate the performance of real- time PCR quantification using both the raw C(t) measurement and the standard...

Ngày tải lên: 09/08/2014, 01:23

... obtained from the patient records at the hospital and the median follow-up for patients still alive was 72.2 months (at the time of these analyses) At the beginning of the project, all the samples ... and bind directly to the cytoplasmic tail of E-cadherin in a mutually exclusive manner α-catenin then links the bound β- or γ-catenin to the actin microfilament network of the cytoskeleton[1,2] ... 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 quantitative...

Ngày tải lên: 09/08/2014, 07:21