Differentiation of pathogenic Bartonella species by infrequent restriction site PCR
Handley, S.A.; Regnery, R.L.
Journal of Clinical Microbiology 38(8): 3010-3015
ISSN/ISBN: 0095-1137 PMID: 10921969 Accession: 003407402
Infrequent restriction site PCR (IRS-PCR) is a recently described DNA fingerprinting technique based on selective amplification of restriction endonuclease-cleaved fragments. Bartonella isolates associated with human disease and related nonhuman isolates were analysed by IRS-PCR genomic fingerprinting. Preparation of DNA templates began with double digestion using 3 different restriction endonuclease combinations. Combinations included the frequently cutting endonuclease HhaI in conjunction with an infrequently cutting endonuclease, EagI, SmaI, or XbaI. Digestion was followed by ligation of oligonucleotide adapters designed with ends complementary to the restriction endonuclease sites.