Weinberg E H, Stakebake J R, Gerone P J
J Bacteriol. 1969 May;98(2):398-402. doi: 10.1128/jb.98.2.398-402.1969.
A plaque technique for the assay of Rickettsia rickettsii is described. The method employs primary chick or green monkey kidney monolayer cell cultures with either an agarose or special Noble agar overlay. Plaques were counted in 6 days and resultant titers correlated well with ld(50) end points obtained by a standard assay in embryonated eggs. Identification of the plaque-forming organisms was accomplished by direct observation of rickettsiae-like bodies in the monolayer lesions, inhibition of plaques by antibiotics, sensitivity of plaques to specific immune serum, and failure to cultivate other microorganisms from the infected cells. Versatility of the test was demonstrated by assaying samples of rickettsiae from several different sources commonly used in our laboratory. These included infected yolk sacs, various cell cultures, and infected guinea pig tissue. Sufficient numbers of viable rickettsiae were present in the cells of a single lesion to permit direct recovery.
本文描述了一种用于检测立氏立克次体的蚀斑技术。该方法采用原代鸡或绿猴肾单层细胞培养物，并覆盖琼脂糖或特制的诺布尔琼脂。在6天内对蚀斑进行计数，所得滴度与通过在鸡胚中进行的标准检测获得的半数致死量（ld50）终点密切相关。通过直接观察单层损伤中类似立克次体的菌体、抗生素对蚀斑的抑制作用、蚀斑对特异性免疫血清的敏感性以及未能从感染细胞中培养出其他微生物来完成对形成蚀斑的生物体的鉴定。通过检测我们实验室常用的几种不同来源的立克次体样本，证明了该检测方法的通用性。这些来源包括感染的卵黄囊、各种细胞培养物以及感染的豚鼠组织。单个损伤的细胞中存在足够数量的活立克次体，以便直接回收。
