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Infection and Immunity, June 2000, p. 3594-3600, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Monitoring Bioluminescent Staphylococcus aureus Infections in Living Mice Using a Novel luxABCDE Construct

Kevin P. Francis,1 Danny Joh,1 Carolyn Bellinger-Kawahara,1 Matthew J. Hawkinson,1 Tony F. Purchio,1 and Pamela R. Contag1,2,*

Xenogen Corporation, Alameda, California 94501,1 and Division of Neonatal and Developmental Medicine, Department of Pediatrics, Stanford University Medical Center, Stanford University, Stanford, California 943052

Received 27 January 2000/Returned for modification 1 March 2000/Accepted 23 March 2000

Strains of Staphylococcus aureus were transformed with plasmid DNA containing a Photorhabdus luminescens lux operon (luxABCDE) that was genetically modified to be functional in both gram-positive and gram-negative bacteria. S. aureus cells containing this novel lux construct, downstream of an appropriate promoter sequence, are highly bioluminescent, allowing the detection of fewer than 100 CFU in vitro (direct detection of exponentially dividing cells in liquid culture). Furthermore, these bacteria produce light stably at 37°C and do not require exogenous aldehyde substrate, thus allowing S. aureus infections in living animals to be monitored by bioluminescence. Two strains of S. aureus 8325-4 that produce high levels of constitutive bioluminescence were injected into the thigh muscles of mice, and the animals were then either treated with the antibiotic amoxicillin or left untreated. Bioluminescence from bacteria present in the thighs of the mice was monitored in vivo over a period of 24 h. The effectiveness of the antibiotic in the treated animals could be measured by a decrease in the light signal. At 8 h, the infection in both groups of treated animals had begun to clear, as judged by a decrease in bioluminescence, and by 24 h no light signal could be detected. In contrast, both groups of untreated mice had strong bioluminescent signals at 24 h. Quantification of CFU from bacteria extracted from the thigh muscles of the mice correlated well with the bioluminescence data. This paper shows for the first time that bioluminescence offers a method for monitoring S. aureus infections in vivo that is sensitive and noninvasive and requires fewer animals than conventional methodologies.

* Corresponding author. Mailing address: Xenogen Corporation, 860 Atlantic Ave., Alameda, CA 94501. Phone: (510) 291-6100. Fax: (510) 291-6196. E-mail: prcontag{at}xenogen.com.

Infection and Immunity, June 2000, p. 3594-3600, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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