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Journal of Clinical Microbiology, March 2001, p. 1097-1104, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.1097-1104.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Antimicrobial Resistance and Bacterial Identification Utilizing a Microelectronic Chip Array

Lorelei Westin,1 Carolyn Miller,2,* Dana Vollmer,2 David Canter,3 Ray Radtkey,3 Michael Nerenberg,3,dagger and James P. O'Connell1

Departments of Advanced Research,1 Assay Development,2 and Molecular Biology,3 Nanogen, Inc., San Diego, California

Received 31 July 2000/Returned for modification 19 September 2000/Accepted 8 December 2000

Species-specific bacterial identification of clinical specimens is often limited to a few species due to the difficulty of performing multiplex reactions. In addition, discrimination of amplicons is time-consuming and laborious, consisting of gel electrophoresis, probe hybridization, or sequencing technology. In order to simplify the process of bacterial identification, we combined anchored in situ amplification on a microelectronic chip array with discrimination and detection on the same platform. Here, we describe the simultaneous amplification and discrimination of six gene sequences which are representative of different bacterial identification assays: Escherichia coli gyrA, Salmonella gyrA, Campylobacter gyrA, E. coli parC, Staphylococcus mecA, and Chlamydia cryptic plasmid. The assay can detect both plasmid and transposon genes and can also discriminate strains carrying antibiotic resistance single-nucleotide polymorphism mutations. Finally, the assay is similarly capable of discriminating between bacterial species through reporter-specific discrimination and allele-specific amplification. Anchored strand displacement amplification allows multiplex amplification and complex genotype discrimination on the same platform. This assay simplifies the bacterial identification process greatly, allowing molecular biology techniques to be performed with minimal processing of samples and practical experience.

* Corresponding author. Mailing address: Assay Development, Nanogen, Inc., 10398 Pacific Center Ct., San Diego, CA 92121. Phone: (858) 410-4718. Fax: (858) 410-4848. E-mail: Cmiller{at}nanogen.com.

dagger Present address: Molecular Reflections, San Diego, Calif.

Journal of Clinical Microbiology, March 2001, p. 1097-1104, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.1097-1104.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.