Isolation and identification procedure for Staphylococcus aureus in laboratory mice and rats by combined use of chromogenic X-SA agar and specific polymerase chain reaction.
Generally, a conventional culture-based examination procedure (detection by egg-yolk salt agar and subsequent identification by phenotypic tests) for confirmation of the presence of S. aureus (SA) in laboratory mice and rats requires approximately 4 days. To improve the culture-based examination procedure for SA in terms of rapidity and reliability, combined use of chromogenic X-SA agar (XSA) and PCR using newly designed specific primers for SA (XSA-PCR) that can shorten the examination time (25.5 hr) was compared with the conventional procedure for SA. In 425 samples from mice and rats, 193 suspected isolates were detected by egg-yolk salt agar (EYSA), and 216 suspected isolates were detected by XSA. In the subsequent identification, 189 of 193 suspected isolates detected by EYSA were identified as SA by phenotypic tests (97.9%), and all 216 suspected isolates detected by XSA were identified as SA by PCR (100%). All SA-positive samples by the conventional procedure were included in the SA-positive samples by XSA-PCR. As a result, XSA-PCR was superior to the conventional procedure in detection rate and identification rate of SA. Therefore, XSA-PCR appears to be an effective tool for examination of SA in laboratory mice and rats that improves precision and shortens the examination time.
Hayashimoto
Nobuhito
N
ICLAS Monitoring Center, Central Institute for Experimental Animals, 1430 Nogawa, Miyamae, Kawasaki, Kanagawa, Japan. nobuh@ciea.or.jp
Goto
Kazuo
K
Takakura
Akira
A
Itoh
Toshio
T
D000362:Agar; D000818:Animals; D000830:Animals, Laboratory / Q000382:microbiology*; D018929:Cell Culture Techniques / Q000379:methods; D002863:Chromogenic Compounds; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification*
D002863:Chromogenic Compounds; D000362:Agar