quantitative polymerase chain reaction using the comparative cq method

... sensitivity of the newly developed method Ten-fold serial dilutions of the extracted viral RNA from the JEV infected culture supernatants were analyzed to define the sensitivity of tests The results ... RD, Watson R, Gelfand DH Detection of specific polymerase chain reaction product by utilizing the 5-3 exonuclease activity of Thermus aquaticus DNA polymerase Proc Natl Acad Sci USA 1991, 88, 72767280 ... Exp Med 1954, 24, 363375 25 Tanaka M Rapid identification of flaviviruses using the polymerase chain reaction J Virol Methods 1993, 41, 311322 26 Warrilow D, Northill JA, Pyke A, Smith GA Single...

Ngày tải lên: 07/08/2014, 18:20

Ngày tải lên: 07/08/2014, 18:21

... was gel purified, the free ends of the plasmid were then shortened using Bal31 nuclease The ends were filled using the Klenow fragment of DNA Polymerase, and then religated The competitor template ... levels and the number of cells expressing these proteins on their surface Given the magnitude of the variations observed by QC-RT-PCR it is unlikely that other methods to quantitate these mRNAs, ... or finger-prick [2] Quantitative methods for reverse transcriptase polymerase chain reaction (RT-PCR) are rapidly surpassing all other methods of quantifying mRNA levels We therefore set out to...

Ngày tải lên: 11/08/2014, 08:20

... of the samples (by either method) These samples were therefore eliminated from the study since they did not meet the definition of sepsis, leaving a total of 400 samples that were evaluated The ... crossing point (Cp) obtained using the LightCycler analysis software v4.05 The Cp represents the point in the amplification cycle where the amplification curve crosses the detection threshold When ... detected in one of these tests, then other samples taken from the infection site were analyzed If this second culture test of the suspected infectious site revealed the presence of the same organism,...

Ngày tải lên: 13/08/2014, 21:21

... Quality of samples submitted METHODOLOGY Samples were processed using either Petroffs’s method or N-acetyl-L-cysteine-sodium hydroxide method 15 A small amount of the processed pellet was used ... hospital patients having a disease other than tuberculosis The latter samples were controls for all the investigations carried out on the test samples Of the 123 samples, 97 samples were taken ... diagnosed as not having tuberculosis, the PCR profile of the individual had changed markedly, suggesting an increase in the bacterial load in the sputum Only seven of the 17 cases i.e those who took...

Ngày tải lên: 06/03/2014, 04:20

... apoptosis and the antitumor immune response of the host These immunized macrophages may sensitize the cytotoxic T lymphocytes of the host, and the sensitized T lymphocytes may attack the residual ... of the patients [23] To clarify this hypothesis, further investigations regarding the correlation between the existence of circulating cancer cells and the host immune response are necessary The ... The reporter dye (6-carboxy-fluorescein: FAM) was covalently attached to the 5’ end of the probe, and the quencher dye (6-carboxy-tetramethyl-rhodamine: TAMRA) was attached to the 3’ end of the...

Ngày tải lên: 18/06/2014, 16:20

... transcriptase polymerase chain reaction The expression levels of the messages in OA and RA are expressed by longitudinal bars relative to those of the control value expressed as 1.0 The lines outside the ... intensity The CT value of the βactin message was used as an internal standard When the target messages were detected in both the control samples and the OA or RA samples by RT-PCR, the ratio for the ... time when the annealed probes were broken by DNA polymerases during the polymerization period The threshold cycle numbers (CT), from which the logarithmic amplification phase of the PCR reaction...

Ngày tải lên: 09/08/2014, 01:24

... differentiate the GPV vaccine strain from the high-virulence strain; nonetheless, we could perform the study on the dynamic distribution of GPV in vivo using these methods, because the animals ... the scope of other diagnostic methods The real-time PCR assay permits the simultaneous detection and quantification of DNA It is useful for understanding the pathogenesis of the disease and the ... (Bio-Rad Corp., Hercules, CA), and the purity was confirmed using the 260/280 nm ratio On the basis of the molecular weight, we calculated the pVP3 copy number using the equations described by Ke...

Ngày tải lên: 12/08/2014, 04:20

... published studies evaluating the use of SepsiTest™ in a clinical setting Key issues in the application of polymerase chain reaction to the diagnosis and management of sepsis The application of PCR ... of pathogenaemia Molecular methods based on polymerase chain reaction (PCR) technology have been developed for infection diagnosis and pathogen identification These methods offer a new approach ... determined by the design of oligonucleotide primers, short pieces of synthesised DNA that bind to either end of the sequence and form the starting point for DNA replication by DNA polymerase For...

Ngày tải lên: 13/08/2014, 16:20

... blood samples In these 59 positive cases, the b3a2 type has the highest rate of 44,07%, followed by the b2a2 type with 32,2%, and the b2a2 + b3a2 combinative type with 23,73% These results indicate ... Trancription -Polymerase Chain Reaction) and Real-Time PCR to detect Philadelphia chromosome containing BCR-ABL fusion gene Results show that the BCR-ABL fusion gene was found in 59 (98,33%) of these ... [2] Khuếch đại týp gene tổ hợp BCR-ABL b2a2 b3a2 kỹ thuật RT-PCR (Reverse Transcription -Polymerase Chain Reaction) sau tiến hành xét nghiệm định lượng Real-Time PCR nhờ 03 nồng độ chuẩn gen β-glucuronidase...

Ngày tải lên: 23/07/2014, 00:21

... left inner ear These findings strongly suggested meningitis with granuloma formation, such as tuberculosis Furthermore, MRI of the lumber vertebrae also suggested the presence of the granulomatous ... (Figure 1) These were confirmed as microcalcification by Von Kossa’s stain Therefore, the PCR assay with toxoplasma-specific primer was performed using nucleic acid extracted from the formalin-fixed ... diagnose the disease definitively Therefore, to make the diagnosis, we employed a PCR assay using nucleic acid extracted from formalin-fixed and paraffin-embedded tissue section of the bone marrow...

Ngày tải lên: 11/08/2014, 03:21

... left inner ear These findings strongly suggested meningitis with granuloma formation, such as tuberculosis Furthermore, MRI of the lumber vertebrae also suggested the presence of the granulomatous ... (Figure 1) These were confirmed as microcalcification by Von Kossa’s stain Therefore, the PCR assay with toxoplasma-specific primer was performed using nucleic acid extracted from the formalin-fixed ... diagnose the disease definitively Therefore, to make the diagnosis, we employed a PCR assay using nucleic acid extracted from formalin-fixed and paraffin-embedded tissue section of the bone marrow...

Ngày tải lên: 11/08/2014, 07:20

... probably due to the reactivation of the latent virus present in them or other invasive procedures that lead to transmission of HCMV In the sub-group III of Group A since the donor’s and the recipient’s ... from the study The mean age of the recipients was 48.5 year and the female to male ratio was 2:3 The age range among recipient males was 23 to 56 and female was 23 to 59 The mean age of the donors ... was copies/ml The highest positive pp65 cell count recorded among them was 78 cells and the lowest was cells In eleven others, the blood sample was positive only by real - time PCR The highest...

Ngày tải lên: 12/08/2014, 02:20

... realtime polymerase chain reaction- based method Hepatology 2007, 46:22-31 Chevaliez S, Bouvier-Alias M, Castéra L, Pawlotsky JM: The Cobas AmpliPrep-Cobas TaqMan real-time polymerase chain reaction ... detect these samples in theory, regardless of the nucleotide polymorphisms Second, the duplex primer/probe assay can estimate the virus levels accurately There are many reports about the underestimation ... detected the HCV 5' UTR sequence Further, both the detection probes were labelled with the same fluorophore, i.e FAM, at the 5' end and with the same quencher dye, i.e Black Hole Quencher (BHQ), at the...

Ngày tải lên: 12/08/2014, 04:20

... to determine the subtypes of dengue virus circulating in the island over the last five years by molecular techniques and so assess the efficacy of this method in the adaptation of the current ... Briefly, RNA was extracted from 280 μl of serum using the QIAamp Viral RNA minikit Reverse transcription and polymerase chain reaction was performed using the One-Step Superscript III/RT/Platinum Taq ... [7,8] The diagnosis of dengue fever by this one-step RT-PCR procedure has reduced the amount of time in getting the results to the patient In comparison with the IgM ELISA or IgG ELISA methods,...

Ngày tải lên: 12/08/2014, 04:21

... marker between leukocytes and the others and D marker between fibroblasts and the others The summary of the individual variations mentioned above is shown in Fig Discussion The AMP PCR is a PCR based ... 4°C Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) The PCR reaction consisted of DNA sample (genomic or digested template), Taq polymerase enzyme (AmpliTaq® Stoffel fragment, ... of the applied enzyme that allows assessing of DNA methylation status of the particular locations throughout the genome The alterations of PCR products in the digested DNA template provide the...

Ngày tải lên: 12/08/2014, 18:22

... Furthermore, the • The rapidity and sensitivity of real-time PCR test results can help the clinician to initiate appropriate cohorting strategies to prevent other critically ill children from acquiring ... and data analysis, and revised the report NJ participated in the design of the study, directed the acquisition and interpretation of clinical data, and revised the report AvL played a substantial ... with the manufacturer's recommended protocol The remaining material was used for real-time PCR testing Real-time PCR Nucleic acids were extracted using the total nucleic acid protocol with the...

Ngày tải lên: 12/08/2014, 23:23

... hospital-acquired infections, not community-acquired For the latter, the rate of initial inadequate treatment typically is lower [4] Furthermore, the daily average treatment cost of community-acquired ... performed in the ICU if the rapidly available PCR results were used to adjust treatments Published interventional data using the new treatment modality are still lacking For the time being, we, therefore, ... interventional studies would allow quantitative observations Besides the cost impact, we analyzed the potential of the new method to lower mortality from sepsis The predicted 2.6% (2.0 to 3.2%)...

Ngày tải lên: 13/08/2014, 21:21

... DANH SÁCH CÁC CHỮ VIẾT TẮT PCR :Polymerase chain reaction DNA :Deoxyrybonucleotide acid TE :Tris EDTA TAE :Tris Acetate EDTA dNTP :Deoxynucleotide triphosphate Taq :Thermus aquaticus UV :Ulta violet ... tính giả  Enzyme DNA polymerase dùng phản ứng PCR phải enzyme chịu nhiệt cao: Taq polymerase đƣợc tách chiết từ vi khuẩn Thermus aquaticus (vi khuẩn suối nƣớc nóng) Taq polymerase cao thấy tƣợng ... PHÁT HIỆN LOÀI NẤM Fusarium spp GÂY BỆNH THỐI XƢƠNG RỒNG (Cactaceae) BẰNG KỸ THUẬT PCR (Polymerase Chain Reaction) Giáo viên hƣớng dẫn KS DƢƠNG THÀNH LAM TS LÊ ĐÌNH DÔN Sinh viên thực PHẠM THỊ...

Ngày tải lên: 17/11/2012, 09:45

... MMC : Microbiological Methods Committee MR : Methyl Red MYP : Mannitol - Egg York - Polymycin NordVal : Nordic System for Validation of Alternative PCR : Polymerase Chain Reaction SE : Relative ... PCR (Polymerase Chain Reaction) Phương pháp PCR phương pháp in vitro để tổng hợp DNA dựa khn trình tự đích DNA ban đầu, khuếch đại, nhân số lượng khn thành hàng triệu nhờ hoạt động enzyme DNA polymerase ... thuật PCR (Polymerase Chain Reaction) PCR phản ứng tổng hợp dây chuyền dùng để khuyếch đại trình tự DNA đích dựa bắt cặp đặc hiệu mồi với DNA mạch khn hoạt tính kéo dài cuả enzyme polymerase...

Ngày tải lên: 16/03/2013, 09:26