恙虫病东方体在无生命培养基内生长的研究

Orientia tsutsugamushi developing through two phases

目的 恙虫病东方体 (Orientiatsutsugamushi;Ot)被认为属专性细胞内寄生。特别是制备大量抗原培养难的问题 ,是近一个世纪以来人们渴望解决的难题。为证实是否能在无生命培养基生长。方法 用Vero -E6个单层组织细胞培养OtJL - 90株与标准Gilliam株 ,在合适时取该培养物接种于Ⅲ号无生命固体培养基培养 ,置 36℃ (± 1℃ )培养 18- 2 4h。结果 可见生长一群无色透明或半透明光滑、湿润、圆形或枫叶样小菌落 ,并产生一种难闻的臭味。两种方法培养物分别做了生物学性状鉴定、形态染色、小白鼠致病力、血清学特异性间接免疫荧光检测。各株Ot经两种培养获得培养物特性基本一致。结论 Ot能在细胞内生长也能在无生命培养基内生长 ,解决了Ot大量抗原培养难的困扰。

Aim Orientia tsutsugamushi (R t) are obligate intracellular parasites,and it is difficult to prepare a fair amount of antigens However,we found that Orientia tsutsugamushi could grow extracellularly well when they cultured with Vero E6 cells,and the results indicated that Orientia tsutsugamushi might grow in abiotic culture mediem Orientia tsutsugamushi cultured by the above method were cultured further in the solid medium for 18～24 hours at 36℃ Small colonies,with achromic transparent,or transulent smooth moist sphere or maple,were observed They generated a kind of stench The appearance of colony of Orientia tsutsugamushi J1-90 strain cultured was similar to that of standard Gilliam strain Orientia tsutsugamushi cultured with two methods are used to identify their biological properties,e g morphology staining,pathogenicity to mice,serological specific detection There were no significant differences in the biological properties of Orientia tsutsugamushi cultured by the two methods The results suggested that Orientia tsutsugamushi might have two development phases