Katayama T, Furuya Y, Yoshida Y, Kaiho I
Kanagawa Prefectural Public Health Laboratory, Japan.
Kansenshogaku Zasshi. 1996 Jun;70(6):561-8. doi: 10.11150/kansenshogakuzasshi1970.70.561.
Primer pairs for PCR were designed from the gene encoding the 17,000-molecular-weight genus-common antigen of Rickettsia japonica, Rickettsia rickettsii, Rickettsia conorii, Rickettsia typhi and Rickettsia prowazekii. Primers R1, R2 were designed for amplifying the genomic DNA from spotted fever group (SFG) rickettsiae and epidemic typhus rickettsiae. Primers Rj5, Rj10 were designed for amplifying the genomic DNA from only R. japonica. Using the primers R1, R2, about a 540-bp fragment was observed by amplifying the genomic DNA from R. japonica, R. rickettsii, R. conorii, Thai tick typhus TT-118, Rickettsia sibirica, Rickettsia montana, Rickettsia askari, R. typhi, R. prowazekii and Katayama strain isolated from the patient infected with SFG rickettsiae. Using the primers Rj5, Rj10, the 357-bp fragment was observed by amplifying the genomic DNA from R. japonica and Katayama strain. Therefore, the Katayama strain was identified to belong to R. japonica. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp bands were amplified from blood of the patients infected with SFG rickettsiae in Kanagawa prefecture. These findings indicate that the causative agent of SFG rickettsiosis in these two patients was R. japonica. The ticks, Ixodes ovatus and Haemaphysalis flava, were collected by out field research in Kanagawa prefecture. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp were amplified from these ticks. This indicates that I. ovatus and H. flava were the vector of R. japonica in Kanagawa prefecture. Also, with the primers R1, R2, about a 540 bp fragment was amplified but with primers Rj5, Rj10, no fragments were amplified from I. ovatus and H. flava. Therefore, these ticks may have SFG rickettsiae other than R. japonica and epidemic typhus rickettsiae.
用于聚合酶链反应(PCR)的引物对是根据编码日本立克次体、立氏立克次体、康氏立克次体、斑疹伤寒立克次体和普氏立克次体17000分子量属共同抗原的基因设计的。引物R1、R2设计用于扩增斑点热群(SFG)立克次体和流行性斑疹伤寒立克次体的基因组DNA。引物Rj5、Rj10设计用于仅扩增日本立克次体的基因组DNA。使用引物R1、R2,通过扩增来自日本立克次体、立氏立克次体、康氏立克次体、泰国蜱传斑疹伤寒TT - 118、西伯利亚立克次体、蒙大拿立克次体、阿斯卡里立克次体、斑疹伤寒立克次体、普氏立克次体以及从感染SFG立克次体的患者中分离出的片山株的基因组DNA,观察到一个约540 bp的片段。使用引物Rj5、Rj10,通过扩增来自日本立克次体和片山株的基因组DNA,观察到357 bp的片段。因此,片山株被鉴定为属于日本立克次体。使用引物R1、R2以及Rj5、Rj10,从神奈川县感染SFG立克次体的患者血液中扩增出了537 bp和357 bp的条带。这些发现表明这两名患者中SFG立克次体病的病原体是日本立克次体。通过在神奈川县的野外研究收集了卵形硬蜱和黄血蜱。使用引物R1、R2以及Rj5、Rj10从这些蜱中扩增出了537 bp和357 bp的片段。这表明卵形硬蜱和黄血蜱是神奈川县日本立克次体的传播媒介。此外,使用引物R1、R2扩增出了一个约540 bp的片段,但使用引物Rj5、Rj10未从卵形硬蜱和黄血蜱中扩增出片段。因此,这些蜱可能携带除日本立克次体和流行性斑疹伤寒立克次体之外的SFG立克次体。