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Specific primer for AP‐PCR identification of Actinobacillus actinomycetemcomitans
Author(s) -
AvilaCampos Mario J.,
Sacchi Cláudio T.,
Whitney Anne M.,
Steigerwalt Arnold G.,
Mayer Leonard W.
Publication year - 1999
Publication title -
journal of clinical periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.456
H-Index - 151
eISSN - 1600-051X
pISSN - 0303-6979
DOI - 10.1034/j.1600-051x.1999.261101.x
Subject(s) - actinobacillus , primer (cosmetics) , amplicon , biology , polymerase chain reaction , genbank , in silico pcr , primer dimer , inverse polymerase chain reaction , genetics , microbiology and biotechnology , multiplex polymerase chain reaction , bacteria , gene , chemistry , organic chemistry
. We used arbitrarily‐primed polymerase chain reaction (AP‐PCR) to design and construct a specific primer pair for the identification of Actinobacillus actinomycetemcomitans . We analyzed 25 DNA samples of A. actinomycetemcomitans isolated from patients with localized juvenile periodontitis. From 90 AP‐PCR primers screened, one amplification product was selected, cloned in pCR II vector, and sequenced. The sequence was used to design a single pair of specific primers. The sequence was compared with GenBank entries using BLAST and showed no significant matches. PCR amplification using the new primer pair AA1416 produced a characteristic 3.5‐Kb band in all A. actinomycetemcomitans DNAs tested. Primer pair AA16S produced no or different amplicon profiles using DNA samples from bacterial species other than A. actino‐mycetemcomitans . Our results show that this single primer pair AA1416 can be used in PCR to identify A. actinomycetemcomitans isolates and differentiate them typing from other periodontal bacteria. These approaches appear promising in facilitating laboratory identification and taxonomy of putative periodontopathogens.