Hirakata Y, Finlay B B, Simpson D A, Kohno S, Kamihira S, Speert D P
Division of Infectious and Immunological Diseases, Department of Pediatrics, University of British Columbia, Vancouver, Canada V5Z 4H4.
J Infect Dis. 2000 Feb;181(2):765-9. doi: 10.1086/315276.
Pseudomonas aeruginosa causes both invasive (bacteremic) and chronic noninvasive infections. A simple in vitro system to screen P. aeruginosa clinical isolates for their capacity to penetrate MDCK cell monolayers has been developed. By means of this system, P. aeruginosa clinical isolates, including 32 blood and 45 respiratory isolates, were examined. When monolayers were infected with 3.5x107 cfu of bacteria, significantly more blood (93.7%) than respiratory (54.4%) isolates (P<.001) were detected in the basolateral medium after 3 h. Penetration ability was usually independent of cytotoxicity. Only 8 (4 blood and 4 respiratory) isolates were cytotoxic, possessed exoU, and passed through the monolayer after epithelial cell death, associated with a marked drop in transepithelial electrical resistance. Conversely, noncytotoxic isolates with high penetration ability but without severe epithelial damage were invasive. This system is well suited for screening clinical isolates and their mutants for specific genes conferring the invasiveness phenotype.
铜绿假单胞菌可引起侵袭性（菌血症性）感染和慢性非侵袭性感染。已开发出一种简单的体外系统，用于筛选铜绿假单胞菌临床分离株穿透MDCK细胞单层的能力。借助该系统，对包括32株血液分离株和45株呼吸道分离株在内的铜绿假单胞菌临床分离株进行了检测。当用3.5×10⁷ cfu细菌感染单层细胞时，3小时后在基底外侧培养基中检测到的血液分离株（93.7%）显著多于呼吸道分离株（54.4%）（P<0.001）。穿透能力通常与细胞毒性无关。只有8株（4株血液分离株和4株呼吸道分离株）具有细胞毒性，携带exoU，且在上皮细胞死亡后穿过单层细胞，同时跨上皮电阻显著下降。相反，具有高穿透能力但无严重上皮损伤的无细胞毒性分离株具有侵袭性。该系统非常适合筛选临床分离株及其突变体中赋予侵袭性表型的特定基因。
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