Departmental Papers (Dental)

Document Type

Journal Article

Date of this Version

12-1993

Publication Source

Molecular Oral Microbiology

Volume

8

Issue

6

Start Page

337

Last Page

343

Abstract

The arbitrary primer polymerase chain reaction (AP-PCR) and Southern blot restriction fragment length polymorphism (RFLP) were used to genotype the periodontal pathogen A. actinomycetemcomitans. Total genomic DNA from 73 strains was extracted by conventional methods. Three random-sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 1% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI, separated on a 0.8% agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterized 5.2 kilobases (kb) DNA fragment cloned from A. actinomycetemcomitans strain Y4. The probe was labeled with digoxigenin, and hybridized fragments were detected with anti-digoxigenin antibody. AP-PCR produced 4–10 DNA bands in the 0.5–5 kb regions and distinguished 9, 13 or 17 genotypes, depending on the specific primer used. Southern blot RFLP analysis revealed 12 hybridization patterns consisting of 1 or 2 DNA fragments (2–23 kb). The addition of the Southern blot analysis to the AP-PCR analysis gave rise to a total of 30 DNA profiles among the 73 A. actinomycetemcomitans study strains. The results indicate that both AP-PCR and Southern blot analysis are useful in clonal analysis of A. actinomycetemcomitans.

Copyright/Permission Statement

This is the peer reviewed version of the following article: [Slots, J., Liu, Y.B., DiRienzo, J.M., Chen, C. (1993). Evaluating two methods for fingerprinting genomes of Actinobacillus actinomycetemcomitans. Molecular Oral Microbiology;8(6):337-43.]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.

Keywords

Actinobacillus actinomycetemcomitans, DNA fingerprinting, polymerase chain reaction, restriction fragment length polymorphism, DNA probe, Southern blot

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Date Posted: 01 March 2022

This document has been peer reviewed.