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Antimicrobial Agents and Chemotherapy, April 2003, p. 1456-1459, Vol. 47, No. 4
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.4.1456-1459.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

PCR-Oligonucleotide Ligation Assay for Detection of Point Mutations Associated with Quinolone Resistance in Streptococcus pneumoniae

Mai-Ha Bui,* Gregory G. Stone,{dagger} Angela M. Nilius, Laurel Almer, and Robert K. Flamm

Infectious Disease Research, Abbott Laboratories, Abbott Park, Illinois

Received 25 February 2002/ Returned for modification 13 October 2002/ Accepted 30 December 2002

We have developed a PCR-oligonucleotide ligation assay to rapidly identify base substitutions in topoisomerase genes that are associated with quinolone resistance in clinical isolates of Streptococcus pneumoniae. Thirty-seven strains for which the ciprofloxacin MICs were >=4 µg/ml and 16 strains for which the MICs were <=2 µg/ml were assayed. Compared with sequence data, the assay correctly identified the DNA bases that encoded amino acids at the four positions most commonly associated with quinolone resistance (Ser79 and Asp83 of ParC and Ser81 and Glu85 of GyrA). Therefore, this procedure can rapidly distinguish single base substitutions associated with quinolone-resistant topoisomerases in S. pneumoniae.

* Corresponding author. Mailing address: Dept. 47T, Bldg. AP52N, Abbott Laboratories, 200 Abbott Park Rd., Abbott Park, IL 60064. Phone: (847) 938-0261. Fax: (847) 935-0400. E-mail: mai.bui{at}abbott.com.

{dagger} Present address: Groton Labs, Pfizer Inc., Groton, CT 06340.

Antimicrobial Agents and Chemotherapy, April 2003, p. 1456-1459, Vol. 47, No. 4
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.4.1456-1459.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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