Emerging Infectious Disease ISSN: 1080-6059

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Human Granulocytic Anaplasmosis, Japan

Norio Ohashi¹

, Gaowa¹, Wuritu, Fumihiko Kawamori, Dongxing Wu, Yuko Yoshikawa, Seizou Chiya, Kazutoshi Fukunaga, Toyohiko Funato, Masaaki Shiojiri², Hideki Nakajima³, Yoshiji Hamauzu, Ai Takano, Hiroki Kawabata, Shuji Ando, and Toshio Kishimoto

Author affiliations: University of Shizuoka and Global Center of Excellence Program, Shizuoka City, Japan (N. Ohashi, Gaowa, Wuritu, F. Kawamori, D. Wu, Y. Yoshikawa); Kochi Institute of Health, Kochi City, Japan (S. Chiya, K. Fukunaga); Shizuoka Institute of Environment and Hygiene, Shizuoka City (F. Kawamori); Muroto Hospital, Muroto, Japan (T. Funato); Kochi Prefectural Aki Hospital, Aki City, Japan (M. Shiojiri, H. Nakajima); Chu-gei Clinic, Aki District, Japan (Y. Hamauzu); National Institute of Infectious Diseases, Tokyo, Japan (A. Takano, H. Kawabata, S. Ando); Okayama Prefectural Institute for Environmental Science and Public Health, Okayama, Japan (T. Kishimoto)

Main Article

Table 1

Results of PCR for select rickettsial organisms for 2 men with human granulocytic anaplasmosis, Kochi Prefecture, Japan*

Days after symptom onset†	Nested PCR result‡
Days after symptom onset†	SFG rickettsia 16S rDNA	Orientia tsutsugamushi 16S rDNA	Anaplasma phagocytophilum p44/msp2	Ehrlichia sp. p28/omp-1
Case-patient 1
3	Negative	Negative	Positive	Negative
19	Negative	Negative	Negative	Negative
Case-patient 2
4	Positive	Negative	Positive	Negative
11	NA	NA	NA	NA

*SFG, spotted fever group; NA, not available.
†After in-hospital treatment with minocycline (200 mg/d), both case-patients improved clinically and were discharged on days 20 and 12, respectively, after symptom onset.
‡Before being used in PCR, blood clots from the patients were homogenized by using BioMasher (Nippi Inc., Tokyo, Japan) and treated overnight with 100 U of streptokinase (WAKO Pure Chemical Industries Ltd, Osaka, Japan). DNA then was extracted by using the QIAamp DNA Mini Kit (QIAGEN, Valencia, CA, USA). Multiplex nested first-step PCR for SFG rickettsiae and O. tsutsugamushi was performed by using the following primers: RO-1F (5'-CCGTAAACGATGAGTGCTAGA-3') and RO-R1 (5'-CCGAGAACGTATTCACCGC-3'). Multiplex nested second-step PCR for SFG rickettsiae 16S rDNA was performed by using the following primers: R-2F (5'-GAAGATTCTCTTTCGGTTTCGC-3') and R-2R (5'-GTCTTGCTTCCCTCTGTAAAC-3'). Multiplex nested second-step PCR for O. tsutsugamushi 16S rDNA was performed by using the following primers: O-2F (5'-GACATGGTAGTCGCGAAAAATG-3') and O-2R (5'-TGCAATCCGAACTGAGATACC-3'). A. phagocytophilum p44/msp2 was amplified by using primers p3726, p4257, p3761, and p4183, and Ehrlichia spp. p28/omp-1 was amplified by using primers conP28-F1, conP28-R1, conP28-F2, and conP28-R2, as described (3,9).

Main Article

¹These authors contributed equally to this article.

²Current affiliation: Ehime Prefectural Central Hospital, Matsuyama, Ehime, Japan.

³Current affiliation: Kochi Medical School, Nankoku, Kochi, Japan.

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