Microbiology Blog: The weblog for microbiologists.

from Christina Egan, Nick M. Cirino and Kimberlee A. Musser in Real-Time PCR: Current Technology and Applications

With the public's reawakened concern regarding use of biological agents as weapons, the rapid detection, discrimination, and identification of pathogenic organisms and toxins has become a priority for state and federal government agencies. High confidence, cost effective, and near real-time diagnostic methods are essential to protecting national health security whether the target is public health, agriculture, commodities, or water supply infrastructures. While culture-based methods have been, and will likely remain, the gold standard for microbiological diagnostics, PCR-based tests offer significant advantages in sensitivity, specificity, speed and data richness that make them invaluable to diagnostic laboratories. We describe the application of real-time PCR methods in biodefense and discuss the use of real-time PCR in biodefense in terms of general workflow and processing considerations, clinical diagnostic applications, environmental diagnostic applications, and multiplex screening. Real-time PCR assays can be either quantitative (qPCR) or qualitative, depending on whether a standard curve is included with the analytical run. Most diagnostic and biodefense applications utilise the qualitative nature of real-time PCR as a detection platform. We consider the future uses and anticipated advances in real-time PCR applications as related to biodefense.

Further reading: Real-Time PCR: Current Technology and Applications

Labels: biodefense, PCR, qPCR, real-time PCR

Following on from their highly acclaimed book "Real-Time PCR: An Essential Guide" published in 2004, Edwards, Logan and Saunders have recently completed a new, fully updated and expanded manual entitled Real-Time PCR: Current Technology and Applications. Publication is planned for January 2009 but the publishers hope to have copies available from October 2008.

Further details: Real-Time PCR: Current Technology and Applications

Labels: PCR, real-time PCR

A number of published reviews on Real-Time PCR have recently been made available as "open-access" papers. These timely and authoritative reviews are written by experts in the field and can be downloaded at: PCR Papers


Further info: PCR Papers

Labels: PCR, real-time PCR

Molecular diagnostic methods are currently being used for the early detection of many viral, bacterial, parasitic and fungal infections. Due to their high specificity and sensitivity, these methods will be inserted in the routine of the clinical laboratories to complement information provided by more conventional methods and above all, to help in the diagnosis of dubious cases. The design of specific probes, coupled to the Polymerase Chain Reaction (PCR) technique provides the required specificity and sensitivity to identify fungal species in a short time. Multicopy genes such as those of ribosomal origin are preferred by many researchers to achieve well defined results. However, because they may generate false positive results, other researchers have looked at single-copy genes of high specificity. This technology is being applied to the identification of frequently found causative agents of systemic mycoses such as Candida albicans, Aspergillus fumigatus and other species within these genera, or agents of less frequent mycoses produced by Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum, Paracoccidioides brasiliensis, among others.

Further reading: Pathogenic Fungi: Insights in Molecular Biology

Labels: diagnosis, fungi, mycology, PCR, real-time PCR