Evaluation of Diagnostic Assay for Rickettsioses Using Duplex Real-Time PCR in Multiple Laboratories in Japan

Japanese journal of infectious diseases

Naota Monma¹  Yukie Shimazu²  Naoya Takamoto³  Asaka Ikegaya⁴  Hiroko Sato⁵  Hiromi Fujita⁶  Seigo Yamamoto⁷  Hiroshi Morita⁸  Fumihiko Kawamori⁹  Yukiko Tamaki^1,10 

[1] Akita Research Center for Public Health and Environment;Department of Food and Nutritional Sciences, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, University of Shizuoka;Department of Virology I, National Institute of Infectious Diseases;Fukushima Institute for Public Health;Hiroshima Prefectural Technology Research Institute, Public Health and Environment Center;Mahara Institute of Medical Acarology;Miyazaki Prefectural Institute for Public Health and Environment;Myojin Clinic;Shizuoka Institute of Environment and Hygiene;Tamaki Hospital

关键词: rickettsiosis;    diagnosis;    Orientia tsutsugamushi;    Rickettsia japonica;    duplex real-time PCR;   

DOI  :  10.7883/yoken.JJID.2017.447

学科分类：传染病学

来源: National Institute of Infectious Diseases

【 摘 要 】

Tsutsugamushi disease and Japanese spotted fever are representative rickettsioses in Japan, and are caused by infection with Orientia tsutsugamushi and Rickettsia japonica, respectively. For molecular-based diagnosis, conventional PCR assays, which independently amplify respective rickettsial DNA, are usually used; however, this approach is time-consuming. Here, we describe a new duplex real-time PCR assay for the simultaneous detection of O. tsutsugamushi and spotted fever group rickettsiae, and its evaluation using several PCR conditions in 6 public health laboratories. The detection limit of the assay was estimated to be 102 copies and the sensitivity was almost identical to that of 3 conventional PCR methods. A total of 317 febrile patients were selected as clinically suspected or confirmed cases of rickettsioses. The detection efficiency of this assay for O. tsutsugamushi from blood or skin (eschar) specimens appeared to be almost the same as that of the conventional PCR method, even when performed in different laboratories, whereas the efficiency for spotted fever group rickettsiae tended to be higher than that of the 2 traditional double PCR assays. Our duplex real-time PCR is thus a powerful tool for the rapid diagnosis of rickettsioses, especially at the acute stage of infection.

【 授权许可】

Unknown