We have evaluated a real-time PCR procedure based on the LightCycler technology for rapid detection of herpes simplex virus (HSV) in genital lesions. Two sets of primers, corresponding to the thymidine kinase and DNA polymerase regions, were used for the amplification reactions in separate capillaries containing the SYBR Green I dye as detection signal. In 28 of 118 samples (24%), HSV was isola...

background aggregatibacter actinomycetemcomitans and tannerella forsythensis are two major pathogens in destructive periodontal disease in humans. the detection of these bacteria is needed for diagnosis and management of the mentioned diseases. objectives we aimed to develop and compare improved multiplex conventional and sybr green real time pcr assays for a specific diagnosis of the organisms...

Doxycycline, a new generation tetracycline antibiotic, is currently one of choice for the treatment and prevention of infections caused by agents of biowarfare. We are developing real time PCR assays to detect tetracycline resistance genes in Gram-negative bacteria. The assay was developed as a multiplex SYBR Green I detection using the Roche Lightcycler and multi-melting peak analysis followed...

The importance of real-time polymerase chain reaction (PCR) has increased steadily in clinical applications over the last decade. Many applications utilize SYBR Green I dye to follow the accumulation of amplicons in real time. SYBR Green I has, however, a number of limitations that include the inhibition of PCR, preferential binding to GC-rich sequences and effects on melting curve analysis. Al...

BACKGROUND Watermelon silver mottle virus (WSMoV), which belongs to the genus Tospovirus, causes significant loss in Cucurbitaceae plants. OBJECTIVES Development of a highly sensitive and reliable detection method for WSMoV. MATERIALS AND METHODS Recombinant plasmids for targeting the sequence of nucleocapsid protein gene of WSMoV were constructed. SYBR Green I real-time PCR was established...

A comparative study was conducted of a novel real-time quantitative PCR test (LightCycler System) with FastStart DNA Master(PLUS) SYBR Green I dye to detect DNA of human herpes virus 6 (HHV-6). Results were compared with those of a real-time quantitative PCR with hybridization probe (HP) formats using the fluorescence resonance energy transfer method, and with those of a single qualitative PCR ...

Real-time polymerase chain reaction (real-time PCR), also known as quantitative PCR, is used to determine relative gene expression or to quantify exact levels of mRNA in cells or tissues. Before the advent of real-time PCR, the major difficulty associated with traditional quantitative or semiquantitative PCR was to ensure that PCR reactions were quantified within the exponential phase of amplif...

A quantitative method based on a real-time PCR assay to enumerate Listeria monocytogenes in biofilms was developed. The specificity for L. monocytogenes of primers targeting the listeriolysin gene was demonstrated using a SYBR Green I real-time PCR assay. The number of L. monocytogenes detected growing in biofilms was 6 x 10(2) CFU/cm2.