Publication: Rapid isolation and susceptibility testing of Leptospira spp. Using a new solid medium, LVW agar
Issued Date
2013-01-01
Resource Type
ISSN
10986596
00664804
00664804
Other identifier(s)
2-s2.0-84872046816
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Antimicrobial Agents and Chemotherapy. Vol.57, No.1 (2013), 297-302
Suggested Citation
Vanaporn Wuthiekanun, Premjit Amornchai, Daniel H. Paris, Sayan Langla, Janjira Thaipadunpanit, Wirongrong Chierakul, Lee D. Smythe, Nicholas J. White, Nicholas P.J. Day, Direk Limmathurotsakul, Sharon J. Peacocka Rapid isolation and susceptibility testing of Leptospira spp. Using a new solid medium, LVW agar. Antimicrobial Agents and Chemotherapy. Vol.57, No.1 (2013), 297-302. doi:10.1128/AAC.01812-12 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/32691
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Rapid isolation and susceptibility testing of Leptospira spp. Using a new solid medium, LVW agar
Abstract
Pathogenic Leptospira spp., the causative agents of leptospirosis, are slow-growing Gram-negative spirochetes. Isolation of Leptospira from clinical samples and testing of antimicrobial susceptibility are difficult and time-consuming. Here, we describe the development of a new solid medium that facilitates more-rapid growth of Leptospira spp. and the use of this medium to evaluate the Etest's performance in determining antimicrobial MICs to drugs in common use for leptospirosis. The medium was developed by evaluating the effects of numerous factors on the growth rate of Leptospira interrogans strain NR-20157. These included the type of base agar, the concentration of rabbit serum (RS), and the concentration and duration of CO2incubation during the initial period of culture. The highest growth rate of NR-20157 was achieved using a Noble agar base supplemented with 10% RS (named LVW agar), with an initial incubation at 30° C in 5% CO2 for 2 days prior to continuous culture in air at 30° C. These conditions were used to develop the Etest for three species, L. interrogans (NR-20161), L. kirschnerii (NR-20327), and L. borgpetersenii (NR-20151). The MICs were read on day 7 for all samples. The Etest was then performed on 109 isolates of pathogenic Leptospira spp. The MIC90values for penicillin G, doxycycline, cefotaxime, ceftriaxone, and chloramphenicol were 0.64 units/ml and 0.19, 0.047, 0.5, and 2 μg/ml, respectively. The use of LVW agar, which enables rapid growth, isolation of single colonies, and simple antimicrobial susceptibility testing for Leptospira spp., provides an opportunity for new areas of fundamental and applied research. Copyright © 2013, American Society for Microbiology. All Rights Reserved.