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Primer Pairs Specific (primer + pair_specific)

Distribution by Scientific Domains
Medical Sciences60%

Selected Abstracts

Simultaneous PCR Detection of the Two Major Bacterial Pathogens of Geranium

JOURNAL OF PHYTOPATHOLOGY, Issue 2 2002
D. L. GLICK
Xanthomonas campestris pv. pelargonii (Xcp) and Ralstonia solanacearum (Rs) are the two most important bacterial pathogens of commercially cultivated geraniums (Pelargonium spp.), both causing bacterial wilt and leaf spot. Asymptomatic infections are important reservoirs of infections in commercial growing facilities. Our objective was to design a multiplex PCR (Polymerase Chain Reaction) assay to detect infection by either or both of these pathogens. We used a previously characterized PCR primer pair for Xcp that amplifies a region of 200 bp. In addition, we designed a new primer pair specific for Rs that amplifies a region of 822 bp. With these two primer pairs, we could detect either or both pathogens. As geranium tissue extracts frequently contain inhibitors of the PCR process, a negative PCR could result from either an accurate indication that the plant was pathogen-free or from a false negative assay. We therefore designed `amplification competence' primers, targeting a portion of the geranium 18 s rRNA gene, and generating a 494-bp amplification product that confirms amplification competence and validates a negative assay result. Thus, the triple primer pair multiplex PCR screens for the two most important bacterial pathogens of geraniums simultaneously confirms amplification competence for each geranium sample. [source]

PCR for the identification of methicillin-resistant Staphylococcus aureus (MRSA) strains using a single primer pair specific for SCCmec elements and the neighbouring chromosome-borne orfX

CLINICAL MICROBIOLOGY AND INFECTION, Issue 10 2005
C. Cuny
Abstract The chromosomal location of the SCCmec elements containing mecA allows the identification of methicillin-resistant Staphylococcus aureus (MRSA) strains by PCR amplification of a sequence covering the right junction of the SCCmec elements and the adjacent chromosomal region encoding the species-specific ORFX. MRSA strains can be identified specifically using one forward primer, with only one or two mismatches, targeting the SCCmec elements of different types, and one reverse primer targeting the orfX region. [source]

A simple method for detecting genetically modified maize in common food products

BIOCHEMISTRY AND MOLECULAR BIOLOGY EDUCATION, Issue 1 2004
Chris Brinegar
Abstract A commercially available leaf DNA extraction and amplification kit has been adapted for the detection of genetically modified material in common food products containing maize. Amplification using published primer pairs specific for the Bacillus thuringiensis delta-endotoxin and maize invertase genes results in a 226-bp invertase PCR product in all samples (an internal positive control) plus a 184-bp product in samples that are genetically modified with the endotoxin gene. The ease and rapidity of DNA extraction and PCR make this exercise especially suitable for advanced-placement high school or lower division college biology students. [source]

The clinical utility of the prostate specific membrane antigen reverse-transcription/polymerase chain reaction to detect circulating prostate cells: an analysis in healthy men and women

BJU INTERNATIONAL, Issue 9 2002
L. Llanes
Objective,To evaluate the overall specificity of nested reverse transcriptase-polymerase chain reaction (RT-PCR) to detect prostate-specific membrane antigen (PSM) mRNA in peripheral blood samples of healthy donors. Subjects and methods,Peripheral blood samples were taken from 60 healthy blood-donors (30 men and 30 women aged < 50 years) and analysed for PSM-mRNA using nested RT-PCR (in ,hot-start' conditions and confirmed using nested EcoRI restriction enzyme). Intron-spanning primer pairs specific for human PSM were deduced from the GenBank sequence (M99487) using gene software. The outer primer pair for PSM was: fwd: 1368 5,-TCACCGGGACTCATGGGTGT-3,; reverse: 1860 5,-GCCTGAAGCAATTCCAAGTCGG-3,. Inner primer pair for PSM was: fwd: 1480 5,-AAGGAAGGGTGGAGACCTAG-3,; reverse: 5-ACTGAACTCTGGGGAAGGAC-3,. The integrity of cDNAs was checked using primer pairs specific for the housekeeping gene ,-actin. The specificity and false-positive rate were calculated assuming that the underlying prostate cancer incidence was nil. Results,The first PCR was negative for all samples (100% specificity; 0% false-positive rate). The nested PCR detected 23 positive samples (23/60, 38%) with an overall specificity of 62% (false positive rate, 38%). Conclusion,Nested RT-PCR of PSM-mRNA in peripheral blood is highly unspecific. Its clinical utility in the management of prostate cancer must be low. Further development is needed of quantitative RT-PCR, primers that identify prostatic PSM or another prostate-specific marker gene to differentiate PSM mRNA from circulating prostate cells and from non-prostatic tissues. [source]