accurate and rapid diagnosis of human cytomegalovirus (hcmv) disease in immunocompromised patients has remained as a challenge. quantitative competitive pcr (qc-pcr) methods for detection of hcmv in these individuals have improved the positive and negative predictive values of pcr for diagnosis of hcmv disease. in this study we used qc-pcr assay, using a co-amplified dna standard, to quantitate...

Understanding biological complexity arising from patterns of gene expression requires accurate and precise measurement of RNA levels across large numbers of genes simultaneously. Real time PCR (RT-PCR) in a microtiter plate is the preferred method for quantitative transcriptional analysis but scaling RT-PCR to higher throughputs in this fluidic format is intrinsically limited by cost and logist...

can assess the safety of biotechnology products. Q-PCR precisely quantifies the amount of the nucleic acid target sequence in DNA or RNA extracted from a variety of samples including animal tissues, final products, cell banks, chromatography eluates, and bulk harvest material. During amplification with target-specific oligonucleotide primers, a fluorogenic oligonucleotide probe — with both repo...

The sensitivity of analysis achievable with PCR has led to the technology being adopted across a range of sectors. For many applications a quantitative result is required, which has driven the development of a range of strategies to determine the amount of starting material in a sample. Approaches such as competitive PCR and limiting dilution analysis have been used as routes to quantification,...

It has long been presumed impossible to measure telomeres in vertebrate DNA by PCR amplification with oligonucleotide primers designed to hybridize to the TTAGGG and CCCTAA repeats, because only primer dimer-derived products are expected. Here we present a primer pair that eliminates this problem, allowing simple and rapid measurement of telomeres in a closed tube, fluorescence-based assay. Thi...

Something I notice often in supervising new graduate students is their frustration with the irreproducibility of their biological data. For example, I am asked frequently ‘‘how many biological replicates do I need for qRT-PCR’’? The answer, of course, lies first with the students themselves. They need as many replicates as will persuade them of the validity of their observations. However, for m...

Real-time PCR methods have become widely used within the past few years. However, real-time PCR is rarely used to study chronic diseases with low pathogen loads, presumably because of insufficient sensitivity. In this report, we developed an integrated nucleic acid isolation and real-time PCR platform that vastly improved the sensitivity of the quantitative detection of the intracellular bacter...

Quantitative real-time PCR (polymerase chain reaction) assays are increasingly used to measure quantities of nucleic acids in samples. They may be used to provide a high-throughput alternative to more traditional biological assays. In this case, a calibration process may be required to convert the PCR measurements onto a more relevant scale. This is most commonly undertaken using simple linear ...

objective: inhibition of apoptosis may favor the onset and progression of cancer. survivin is an inhibitor of apoptosis that has been considered as a potential marker for diagnostic and/or prognostic of bladder cancer. the survivin protein regulates both cell division and cell death and is overexpressed in the vast majority of human cancers. in this study, the expression pattern and potential p...