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Dot-ELISA Rapid Test Using Recombinant 56-kDa Protein Antigens for Serodiagnosis of Scrub Typhus

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  • Armed Forces Research Institute of Medical Science (AFRIMS), Royal Thai Army, Bangkok, Thailand; Naval Medical Research Center (NMRC), Silver Spring, Maryland; Preventive Medicine and Biometrics Department, Uniformed Services University of the Health Sciences (USUHS), Bethesda, Maryland

We developed a rapid dot–enzyme-linked immunosorbent assay (dot-ELISA) using the combination of recombinant 56-kDa protein antigens that exhibited broad reactivity with serum antibodies against the four most prevalent strains (Karp, Kato, Gilliam, and TA763) of Orientia tsutsugamushi. The assay is rapid (30 minutes), and can be done at room temperature, and results can be read by the naked eye. Only a simple shaker is required to wash the membrane. Sera from 338 patients suspected of being ill with scrub typhus from rural hospitals around Thailand were tested using this dot-ELISA. Seventy-five (22.2%) patients were found to be positive. The sensitivity and specificity of dot-ELISA were determined using the indirect immunofluorescent assay (IFA) test as the gold standard, with the cutoff titer of immunoglobulin peroxidase conjugate M (IgM)/G (IgG) greater than 1:400/1:400. The dot-ELISA had a sensitivity of 98.5%, a specificity of 96.3%, a positive predictive value of 86.7%, and a negative predictive value of 99.6% for the acute-phase specimens. The results indicate that dot-ELISA rapid test using recombinant 56-kDa protein antigen was comparable with the IFA test and may be very useful for the diagnosis of scrub typhus in rural hospitals, where IFA is not available.

Author Notes

* Address correspondence to Wei-Mei Ching, Viral and Rickettsial Disease Program, Infectious Diseases Department, Code 41, Naval Medical Research Center, Silver Spring, MD 20910-7500. E-mail: wei.ching@med.navy.mil

Financial support: Support for this study was provided by the Royal Thai Army (to W.R.). Part of the work collaborating with the Naval Medical Research Center (NMRC) was supported by Work Unit Number 6000 (RAD1.J.A0310).

Authors' addresses: Wuttikon Rodkvamtook, Dharadhida Bodhidatta, Jariyanart Gaywee, Narongrid Sirisopana, and Manerat Kityapan, Microbiology, Armed Forces Research Institute of Medical Science, Bangkok, Thailand, E-mails: ltwutti@hotmail.com, dbodhi@hotmail.com, jariyanart@yahoo.com, Narongrid1956@yahoo.com, and krataii_usagi@hotmail.com. Zhiwen Zhang, Chien-Chung Chao, Erin Huber, and Wei-Mei Ching, Viral and Rickettsial Diseases Department, Naval Medical Research Center, Silver Spring, MD, and Preventive Medicine and Biometrics Department, Uniformed Services University of the Health Sciences, Bethesda, MD, E-mails: Zhiwen.zhang@med.navy.mil, chien-chung.chao@med.navy.mil, erin.huber@med.navy.mil, and wei.ching@med.navy.mil. John Grieco and Michael Lewis, Preventive Medicine and Biometrics Department, Uniformed Services University of the Health Sciences, Bethesda, MD, E-mail: johngrieco88@gmail.com and dr.michael.lewis@gmail.com.

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