Jpn. J. Infect. Dis., 57, 103-106, 2004

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Original Article

Factors Improving the Propagation of Simkania negevensis Strain Z in Cell Culture

Tetsuya Yamaguchi^1,2* Tsutomu Yamazaki^2,3,4 Miyuki Inoue³, Motohiko Ogawa², Sadashi Shiga², Yuichi Nakagawa¹, Toshio Kishimoto²,　Kazunobu Ouchi⁵ and Takehiko Ohzeki¹

¹Department of Pediatrics, Hamamatsu University School of Medicine, Shizuoka 431-3192, ²Laboratory of Rickettsia and Chlamydia, Department of Virology I, National Institute of Infectious Diseases, Tokyo 162-8640, ³Department of Pediatrics and ⁴Department of Infectious Diseases and Infection Control, Saitama Medical School, Saitama 350-0495 and ⁵Department of Pediatrics, Kawasaki Medical School, Okayama 701-0192, Japan

(Received February 5, 2004. Accepted March 11, 2004)

*Corresponding author: Mailing address: Department of Pediatrics, Hamamatsu University School of Medicine, Handayama 1-20-1, Hamamatsu, Shizuoka 431-3192, Japan. Tel: +81-53-435-2312, Fax: +81-53-435-2311, E-mail: tetsuyaf@hama-med.ac.jp

SUMMARY: The purpose of the present study was to develop an optimal method for culturing Simkania negevensis. Centrifugation was effective for the propagation of S. negevensis, but sonication was not effective. The addition of cycloheximide to the culture medium significantly decreased the number of inclusions. Pretreatment of host monolayers with diethylaminoethyl-dextran or polyethylene glycol was detrimental. The most optimal conditions were centrifugation of the inocula onto untreated Vero cells, and culture in RPMI 1640 medium containing 10% fetal calf serum without cycloheximide or antimicrobial agents.

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