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Quantitative real‐time PCR based on single copy gene sequence for detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis by Juan M Morillo; Laura Lau; Mariano Sanz; David Herrera; Augusto Silva is a Dentistry article available to read on EtoBox.

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**Objective:** To establish a method for quantification of __Actinobacillus actinomycetemcomitans__ and __Porphyromonas gingivalis__ from subgingival plaque by real‐time polymerase chain reaction (PCR) technique. **Material and methods:** Bacterial cells from both species were obtained from type culture and counted microscopically. Cellular suspension in sterile destilled water was used for DNA extraction by boiling for 20 min, with a mineral oil cover. Primers for PCR were selected from sequences of LktC gene __(A. actinomycetemcomitans)__ and Arg‐gingipain __(P. gingivalis)__ to yield amplicons below 100 bp. SYBR Green I based real‐time PCR was adjusted to quantify separately both species. **Results:** A good sensitivity and specificity were obtained for both species, although the yield was better for __A. actinomycetemcomitans__. A good repeatibility of cycle threshold (__C__~T~) was encountered, so coefficient of variation was below 6% at every initial copy number. **Conclusion:** A new method of quantification of __A. actinomycetemcomitans__ and __P. gingivalis__ based on SYBR Green real‐time PCR is presented. Its good sensibility and repeatibility will allow its application t

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Author

Juan M Morillo; Laura Lau; Mariano Sanz; David Herrera; Augusto Silva

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0022-3484)

Published

2003

Language

EN

Field

Dentistry (Health Sciences)