16S rDNA PCR-denaturing gradient gel electrophoresis in detennining proportions of coexisting Actinobacillus actinomycetemcomitans strains

: Ihalin R, Asikainen S

Publisher: ELSEVIER SCIENCE BV

: 2006

: Journal of Microbiological Methods

: JOURNAL OF MICROBIOLOGICAL METHODS

: J MICROBIOL METH

: 65

: 3

: 417

: 424

: 8

: 0167-7012

DOI: https://doi.org/10.1016/j.mimet.2005.08.016

: https://www.sciencedirect.com/science/article/pii/S0167701205002769?via=ihub

Certain serotypes of Actinobacillus actinomycetemcomitans seem to prefer coexistence in vivo. The 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) was tested for its capability to distinguish coexisting A. actinomycetemcomitans strains of different serotypes or genetic lineages and to determine their proportions in vitro. The migration pattern of the PCR amplicon from serotype c differed from those of the other serotypes. Contrary to the strains of serotypes c, d, and e, strains of serotypes a, b, and f consistently demonstrated intra-serotype migration patterns similar to each other. Since the migration patterns differed between serotype c and b strains a strain of each was used to determine their proportional representation in a strain mixture. The strains were distinguishable from each other above the 5% PCR-DGGE detection level (12.5 ng DNA/1.5 x 10(6) cells). DGGE provides a promising tool for in vitro studies on the coexistence of different genetic lineages of A. actinomycetemcomitans. (c) 2005 Elsevier B.V. All rights reserved.