Rapid quantification of periodontitis-related bacteria using a novel modification of Invader PLUS technologies

The Invader PLUS technology is a sensitive, rapid method for the detection and quantification of nucleic acid. While the original technology is based on the amplification by polymerase chain reaction (PCR) of the target sequence followed by its detection using the Invader technology, the current mod...

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Bibliographic Details
Published in:Microbiological research 2010-01, Vol.165 (1), p.43-49
Main Authors: Tadokoro, Kenichi, Yamaguchi, Toshikazu, Kawamura, Katsumi, Shimizu, Hajime, Egashira, Toru, Minabe, Masato, Yoshino, Toshiaki, Oguchi, Hirokazu
Format: Article
Language:English
Subjects:
Actinobacillus actinomycetemcomitans
Bacteria - chemistry
Bacteria - genetics
Bacteria - isolation & purification
DNA, Bacterial - analysis
DNA, Bacterial - genetics
Fusobacterium nucleatum
Genetic Techniques
Humans
Invader
Periodontitis
Periodontitis - microbiology
Porphyromonas gingivalis
Prevotella intermedia
Quantification
Real-time PCR
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Summary:The Invader PLUS technology is a sensitive, rapid method for the detection and quantification of nucleic acid. While the original technology is based on the amplification by polymerase chain reaction (PCR) of the target sequence followed by its detection using the Invader technology, the current modification allows simultaneous PCR amplification and Invader reaction. The PCR primers and the Invader probes are designed to operate at the same temperature. This allows simpler design and faster results. This technology has been applied for the quantification of six periodontitis-related bacteria ( Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Toreponema denticola, Tannerella forsythensis and Fusobacterium nucleatum). Direct comparison of this modified Invader PLUS with real-time PCR demonstrated similar linear range. Furthermore, testing of 64 volunteers showed a good correlation between both technologies with correlation factors r2 spanning between 0.827 and 0.987. We demonstrated here that the proposed improvement of the Invader PLUS allows the detection and quantification of DNA sequences using a simple design and protocol that can be implemented in clinical testing.
ISSN:0944-5013
1618-0623
DOI:10.1016/j.micres.2008.06.001