Evaluation of Enzyme-Linked Immunosorbent Assay Using Recombinant 56-kDa Type-Specific Antigens Derived from Multiple Orientia tsutsugamushi Strains for Detection of Scrub Typhus Infection

Su-Lin Yang Center for Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Republic of China;

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Kun-Hsien Tsai Department of Public Health and Institute of Environmental Health, College of Public Health, National Taiwan University, Taipei, Republic of China

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Hsiang-Fei Chen Center for Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Republic of China;

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Jun-Yu Luo Center for Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Republic of China;

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Pei-Yun Shu Center for Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Republic of China;

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Scrub typhus is caused by the intracellular bacterium Orientia tsutsugamushi. The 56-kDa type-specific antigen (TSA) displays a significant antigenic variation across different O. tsutsugamushi strains. To minimize the influence of the antigenic diversity of TSA on assay sensitivity, we developed a mixed-TSA enzyme-linked immunosorbent assay (mixed-TSA ELISA) using a mixture of recombinant TSAs of prototype (Karp, Gilliam, and Kato) and local (TW-1, TW-10, TW-19, and TW-22) O. tsutsugamushi strains as antigens to detect immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies against O. tsutsugamushi. These four local strains covered a major part of the total genetic diversity of TSA gene of O. tsutsugamushi in Taiwan. A total of 109 acute-phase serum samples from O. tsutsugamushi polymerase chain reaction–positive, scrub typhus patients, and 82 negative control serum samples from non-scrub typhus cases were used for evaluation of the recombinant TSA-based ELISA. We compared the performance of the mixed-TSA ELISA with immunofluorescence assay (IFA), which is considered the gold standard method for the serological diagnosis of scrub typhus. The results indicated that the sensitivity of IgM mixed-TSA ELISA (80.7%) was significantly higher than that of IgM IFA (68.8%). We demonstrated that the mixed-TSA ELISA had a high sensitivity and specificity and can be used for screening of scrub typhus patient in the early phase of the disease.

Author Notes

Address correspondence to Pei-Yun Shu, Center for Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, No. 161, Kunyang St., Taipei 11561, Republic of China. E-mail: pyshu@cdc.gov.tw

Conflicts of Interest: PYS and SLY have a patent detection kit for diagnosis of scrub typhus and detection method thereof pending.

Financial support: This work was supported in part by grants MOHW104-CDC-C-315-000115 and MOHW105-CDC-C-315-123504 from Centers for Disease Control, Ministry of Health and Welfare, Taiwan, Republic of China.

Authors’ addresses: Su-Lin Yang, Hsiang-Fei Chen, Jun-Yu Luo, and Pei-Yun Shu, Center for Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Republic of China, E-mails: cerline@cdc.gov.tw, xiangfei@cdc.gov.tw, lions8022@cdc.gov.tw, and pyshu@cdc.gov.tw. Kun-Hsien Tsai, Department of Public Health and Institute of Environmental Health, College of Public Health, National Taiwan University, Taipei, Republic of China, E-mail: kunhtsai@ntu.edu.tw.

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