Immunomagnetic PCR\ DNA Probe\ Porphyromonas gingivalis

Christian Mouton Christian.Mouton at greb.ulaval.ca
Fri Sep 15 15:59:14 EST 1995


The following is the summary of a paper which has been accepted for
publication by J. Clin. Microbiol.

Of particular interest is the fact that the DNA probe  and  the PCR
protocol specific for Porphyromonas gingivalis were derived from the
sequence of a random amplified polymorphic DNA (RAPD) species-specific
fragment. We are currently developing  similar procedures for the diagnosis
of other fastidious gram negative anaerobes in mixed infections.

Immunomagnetic PCR and a DNA Probe for the Detection and Identification of
Porphyromonas gingivalis.

Riad M. Benkirane,  Emmanuelle Guillot, and Christian Mouton

The aim of this study that we describe was to combine an immunomagnetic
separation and a polymerase chain reaction followed by dot blot
hybridization with a DNA probe for the detection and identification of
Porphyromonas gingivalis. Immunomagnetic particles were coated with
monoclonal antibody specific for P. gingivalis and incubated with a
suspension of seven oral bacterial species spiked with various dilutions of
P. gingivalis. Beads with their load of bound bacteria were boiled in
water, and the target DNA in the supernatant was amplified with a primer
pair to generate a 593-bp PCR fragment specific for P. gingivalis. Finally,
the product of amplification was detected by dot-blot hybridization with a
digoxigenin-labeled 593-bp probe. The detection limit was determined at 100
bacterial cells/ml. The immunomagnetic-PCR/DNA probe procedure described
here should be useful for the rapid, specific and sensitive detection and
identification of P. gingivalis in clinical samples.


-----------------------------Christian Mouton-----------------------------
Groupe de Recherche en Écologie Buccale
tél. (418)656-5872
Faculté de médecine dentaire
fax. (418)656-2861
Université Laval
Email: Christian.Mouton at greb.ulaval.ca
Québec (Québec)
G1K 7P4  CANADA








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