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Clinical and Vaccine Immunology, February 2007, p. 173-181, Vol. 14, No. 2
1071-412X/07/$08.00+0     doi:10.1128/CVI.00347-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Development of Serological Assays for Thottapalayam Virus, an Insectivore-Borne Hantavirus{triangledown}

Megumi Okumura,1 Kumiko Yoshimatsu,1 Sanit Kumperasart,2 Ichiro Nakamura,3 Michiko Ogino,1 Midori Taruishi,1 Araya Sungdee,2 Sirima Pattamadilok,2 Ima Nurisa Ibrahim,4 Sri Erlina,4 Takashi Agui,5 Richard Yanagihara,6 and Jiro Arikawa1*

Institute for Animal Experimentation, Graduate School of Medicine, Hokkaido University, Sapporo 060-8638, Japan,1 National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi 11000, Thailand,2 Hokkaido University Research Center for Zoonosis Control, Sapporo 060-0818, Japan,3 National Institute of Health Research and Development, Jakarta, Indonesia,4 Laboratory of Experimental Animal Science, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan,5 Department of Pediatrics, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, Hawaii 968136

Received 19 September 2006/ Returned for modification 30 October 2006/ Accepted 29 November 2006

Thottapalayam virus (TPMV), a member of the genus Hantavirus in the family Bunyaviridae, was isolated from an insectivore, Suncus murinus (musk shrew), captured in southern India in 1964. While the isolation of TPMV predates the discovery of the prototype Hantaan virus, little is known about its genetics and biology. To date, preliminary evidence suggests that TPMV differs significantly, both antigenically and genetically, from all known rodent-borne hantaviruses. However, since detailed epizootiological studies have not been conducted, it is unclear if TPMV is naturally harbored by an insectivore host or if TPMV represents a "spillover" from its natural rodent reservoir host. Moreover, to what extent TPMV causes infection and/or disease in humans is not known. To address these issues, we first studied the antigenic profile of TPMV using monoclonal antibodies against Hantaan and Seoul viruses and polyclonal immune sera against Puumala virus and TPMV. Armed with this newfound information, we developed an enzyme-linked immunosorbent assay system for the diagnosis of TPMV infections in shrews and humans, using a recombinant TPMV N antigen manipulated to have an E5/G6 epitope to be captured by monoclonal antibody clone E5/G6. Using this assay, we found anti-TPMV antibodies in sera from a patient with high fever of unknown etiology in Thailand and from two shrews captured in Indonesia. Seropositivity was verified by the indirect immunofluorescence antibody test, Western blotting analysis, and focus reduction neutralization test. Collectively, our data indicate that TPMV is harbored by Suncus murinus as its host in nature and is capable of infecting humans.

* Corresponding author. Mailing address: Institute for Animal Experimentation, Hokkaido University Graduate School of Medicine, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, Japan. Phone: 81-11-706-6905. Fax: 81-11-706-7879. E-mail: j_arika{at}med.hokudai.ac.jp.

{triangledown} Published ahead of print on 20 December 2006.

Clinical and Vaccine Immunology, February 2007, p. 173-181, Vol. 14, No. 2
1071-412X/07/$08.00+0     doi:10.1128/CVI.00347-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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