Development of a Real-Time Fluorescence PCR Assay for Rapid Detection of the Diphtheria Toxin Gene
AUTOR(ES)
Mothershed, Elizabeth A.
FONTE
American Society for Microbiology
RESUMO
We developed and evaluated a real-time fluorescence PCR assay for detecting the A and B subunits of diphtheria toxin (tox) gene. When 23 toxigenic Corynebacterium diphtheriae strains, 9 nontoxigenic C. diphtheriae strains, and 44 strains representing the diversity of pathogens and normal respiratory flora were tested, this real-time PCR assay exhibited 100% sensitivity and specificity. It allowed for the detection of both subunits of the tox gene at 750 times greater sensitivity (2 CFU) than the standard PCR (1,500 CFU). When used directly on specimens collected from patients with clinical diphtheria, one or both subunits of the tox gene were detected in 34 of 36 specimens by using the real-time PCR assay; only 9 specimens were found to be positive by standard PCR. Reamplification by standard PCR and DNA sequencing of the amplification product confirmed all real-time PCR tox-positive reactions. This real-time PCR format is a more sensitive and rapid alternative to standard PCR for detection of the tox gene in clinical material.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=154649Documentos Relacionados
- Development of a Real-Time PCR Assay for Rapid Detection and Quantification of Alexandrium minutum (a Dinoflagellate)
- Duplex Real-Time PCR Assay for Rapid Detection of Ampicillin-Resistant Enterococcus faecium
- Real-Time Fluorescence-Based PCR for Detection of Malaria Parasites
- Development of a Real-Time PCR Assay for Quantitative Detection of Encephalitozoon intestinalis DNA
- Detection of Staphylococcus aureus Enterotoxins A to D by Real-Time Fluorescence PCR Assay
