The rapid identification of bacteria and fungi directly from the blood of patients with suspected bloodstream infections aids in diagnosis and guides treatment decisions. The development of an automated, rapid, and sensitive molecular technology capable of detecting the diverse agents of such infections at low titers has been challenging, due in part to the high background of genomic DNA in blo...

Rapid pathogen testing is vital to the food industry. Enzyme immunoassays (EIA) provide reliable negative results in 48 h, but a presumptive positive (suspect) EIA result must be confirmed by traditional culture methods, requiring an additional 72 h. Polymerase chain reaction (PCR) testing technology is accepted as an accurate diagnostic tool. However, traditional PCR techniques can require sev...

The polymerase chain reaction (PCR) is an easy, economic, convenient and biochemical technology in molecular biology for the amplification of thousands to millions of copies of a particular DNA sequence. The method consists of repetitive cycles of three steps: denaturation, hybridization, and polymerase extension, that raising temperature to break hydrogen bonds for formation of the single stra...

Allele-specific polymerase chain reaction (AS-PCR) is a rapid and cost-effective single nucleotide polymorphism (SNP) genotyping method compared to multiplex or real-time PCR. The SNPs occurring in mitochondrial DNA (mtSNPs) the most abundant humans can be used human identification involving mass fatality incidents. Nevertheless, application of AS-PCR has yet widely applicable forensic investig...

Nucleic acid amplification can provide sensitive and specific molecular detection and quantification. Since its introduction PCR has been the dominant amplification technology; its widespread use has stimulated improvements in enzymes, instrumentation, and analytical methods. Initial use of PCR was largely qualitative: positive target detection, or inference of target absence above a detection ...

Center for Environmental Health Sciences and Division of Toxicology, Whitaker College of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 The efficacy of PCR is measured by its specificity, efficiency (i.e. yield), and fidelity. A highly specific PCR will generate one and only one amplification product that is the intended target sequence. Mo...

Center for Environmental Health Sciences and Division of Toxicology, Whitaker College of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 The efficacy of PCR is measured by its specificity, efficiency (i.e. yield), and fidelity. A highly specific PCR will generate one and only one amplification product that is the intended target sequence. Mo...

Overlapping PCR is routinely used in a wide number of molecular applications. These include stitching PCR fragments together, generating fluorescent transcriptional and translational fusions, inserting mutations, making deletions, and PCR cloning. Overlapping PCR is also used for genotyping by traditional PCR techniques and in detection experiments using techniques such as loop-mediated isother...

The Seeplex TB Detection-2 assay (Rockville, MD) is a nested endpoint PCR for the Mycobacterium tuberculosis complex (MTBC) targets IS6110 and MPB64 that utilizes dual priming oligonucleotide technology. When used to detect the presence of MTBC DNA in formalin-fixed paraffin-embedded tissue specimens, the sensitivity and specificity of this assay is equivalent to a labor-intensive traditional e...

DNA methylation of CpG sites in promoter regions of several cancer related genes, such as O6-MGMT, hMLH1, p14ARF, p16INK4a, RASSF1A and APC1A, has been actively explored recently. Much of the data has been obtained using a variation of an allele-specific PCR assay known as methylation specific PCR. This technique is objectionable for a number of methodological limitations and drawbacks. We want...