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Volume 28, Number 11—November 2022
Research Letter

Serologic Evidence of Human Exposure to Ehrlichiosis Agents in Japan

Hongru Su, Kenji Kubo, Shigetoshi Sakabe, Shinsuke Mizuno, Nobuhiro Komiya, Shigehiro Akachi, Hiromi Fujita1, Kozue Sato, Hiroki Kawabata, Hiromi Nagaoka, Shuji Ando, and Norio OhashiComments to Author 
Author affiliations: University of Shizuoka, Shizuoka, Japan (H. Su, N. Ohashi); Japanese Red Cross Wakayama Medical Center, Wakayama, Japan (K. Kubo, S. Mizuno, N. Komiya); Ise Red Cross Hospital, Ise, Japan (S. Sakabe); Mie Prefecture Health and Environment Research Institute, Yokkaichi, Japan (S. Akachi); Mahara Institute of Medical Acarology, Anan, Japan (H. Fujita); National Institute of Infectious Diseases, Tokyo, Japan (K. Sato, H. Kawabata, S. Ando); Shizuoka Institute of Environment and Hygiene, Shizuoka (H. Nagaoka)

Main Article

Figure

Western blots using serum samples from a febrile patient (case 1) in Wakayama Prefecture in study showing serologic evidence of human exposure to ehrlichiosis agents in Japan. Serum samples were collected from the patient on day 2 and 17 after onset of illness. Human THP-1 and canine DH82 cells were uninfected or infected with Ehrlichia chaffeensis . THP-1 cells were also infected with Anaplasma phagocytophilum. Cell lysates were separated and Western blot was performed as described (Appendix). We used uninfected THP-1 and DH82 cells as negative lysate controls. We used rabbit serum against recombinant P44 antigens specific for A. phagocytophilum and recombinant P28 antigens specific for E. chaffeensis (1:10,000 dilution) as positive serum controls. We used serum from a healthy donor as a negative control serum (Precision for Medicine, https://www.precisionbiospecimens.com). The patient’s serum samples and negative control serum were diluted 1:250 and used to probe the blots. We used alkaline-phosphatase-conjugated goat anti-human IgM μ-chain and anti-human IgG γ-chain (Thermo Fisher Scientific, https://www.thermofisher.com) as secondary antibodies. Arrows indicate E. chaffeensis-specific P28 antigens (encoded by a p28 multigene family). Arrowhead shows A. phagocytophilum-specific P44 antigen (encoded by a p44 multigene family).

Figure. Western blots using serum samples from a febrile patient (case 1) in Wakayama Prefecture in study showing serologic evidence of human exposure to ehrlichiosis agents in Japan. Serum samples were collected from the patient on day 2 and 17 after onset of illness. Human THP-1 and canine DH82 cells were uninfected or infected with Ehrlichia chaffeensis . THP-1 cells were also infected with Anaplasma phagocytophilum. Cell lysates were separated and Western blot was performed as described (Appendix). We used uninfected THP-1 and DH82 cells as negative lysate controls. We used rabbit serum against recombinant P44 antigens specific for A. phagocytophilum and recombinant P28 antigens specific for E. chaffeensis (1:10,000 dilution) as positive serum controls. We used serum from a healthy donor as a negative control serum (Precision for Medicine, https://www.precisionbiospecimens.com). The patient’s serum samples and negative control serum were diluted 1:250 and used to probe the blots. We used alkaline-phosphatase-conjugated goat anti-human IgM μ-chain and anti-human IgG γ-chain (Thermo Fisher Scientific, https://www.thermofisher.com) as secondary antibodies. Arrows indicate E. chaffeensis-specific P28 antigens (encoded by a p28 multigene family). Arrowhead shows A. phagocytophilum-specific P44 antigen (encoded by a p44 multigene family).

Main Article

1Current affiliation: Northern Fukushima Medical Center, Date, Japan.

Page created: September 08, 2022
Page updated: October 24, 2022
Page reviewed: October 24, 2022
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