This study compared SYBR Green real-time quantitative PCR (qPCR) with standard plate counting for the enumeration of Streptococcus mutans in oral samples. Oral samples (n = 710) were collected from high-caries-risk children for quantification of S. mutans by qPCR using primer pairs. The S. mutans copy number was calculated with reference to a qPCR quantification cycle (Cq) standard curve and co...

Real-time quantitative PCR (qPCR) assay of sputum or nasopharyngeal specimens has shown promising results in the detection of pneumococcal community-acquired pneumonia (PncCAP). We applied qPCR for the autolysin gene (lytA) and compared sputum and nasopharyngeal swab (NPS) pneumococcal loads in elderly patients with community-acquired pneumonia (CAP), and specifically in patients with PncCAP, t...

Following its invention 25 years ago, PCR has been adapted for numerous molecular biology applications. Gene expression analysis by reverse-transcription quantitative PCR (RT-qPCR) has been a key enabling technology of the post-genome era. Since the founding of BioTechniques, this journal has been a resource for the improvements in qPCR technology, experimental design, and data analysis. qPCR a...

Quantitative PCR (qPCR) is a commonly used molecular biology technique that enables the precise determination of the amount of DNA or RNA in a sample of interest. By measuring the amount of DNA present at each cycle of a PCR, the concentration in an unknown sample can be calculated by comparison to a set of known standard samples. This real-time measurement of the PCR product is achieved using ...

Development of rapid and sensitive detection methods for group B streptococci (GBS) in pregnant women remains useful in order to adequately identify pregnant women at risk of transferring GBS to their neonate. This study compared the CDC recommended sampling and culture method with two qPCR methods for detecting GBS colonization. For a total of 100 pregnant women at 35-37 weeks of gestation, on...

BACKGROUND Improved understanding of Neisseria meningitidis (Nm) carriage biology and better methods for detection and quantification would facilitate studies of potential impact of new vaccines on colonization and transmission in adolescents. METHODS We performed plate cultures on 107 oropharyngeal swabs stored frozen in skim milk tryptone glucose glycerol (STGG) broth and previously positiv...

Scientific conferences fulfil many roles, but one of the most mportant ones is that they help shape the direction in which a scintific discipline grows by promoting person-to-person exchanges f information, ideas and constructive criticisms between scientists rom different backgrounds. This interaction also helps to idenify areas of controversy and promotes efforts to address and, it is oped, r...

The polymerase chain reaction (PCR) is usually described as a simple, sensitive and rapid technique that uses oligonucleotide primers, dNTPs and a heat stable Taq polymerase to amplify DNA. It was invented by Mullis and co-workers [1,2] in the early eighties, who were awarded the 1993 Nobel Prize for chemistry for this discovery. With the discovery of real-time PCR in the nineties the method to...

In microbial food safety, molecular methods such as quantitative PCR (qPCR) and next-generation sequencing (NGS) of bacterial isolates can potentially be replaced by diagnostic shotgun metagenomics. However, the methods for pre-analytical sample preparation are often optimized for qPCR, and do not necessarily perform equally well for qPCR and sequencing. The present study investigates, through ...

Quantitative real-time PCR (qPCR) is a sensitive technique for the detection and quantitation of specific DNA sequences. Here we describe a Taqman qPCR assay for quantification of tissue-localized, adoptively transferred enhanced green fluorescent protein (EGFP)-transgenic cells. A standard curve constructed from serial dilutions of a plasmid containing the EGFP transgene was (i) highly reprodu...