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Molecular Detection and Typing of Dengue Viruses from Archived Tissues of Fatal Cases by RT-PCR and Sequencing: Diagnostic and Epidemiologic Implications

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  • Infectious Diseases Pathology Branch, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia
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Diagnosis of dengue virus (DENV) infection in fatal cases is challenging because of the frequent unavailability of blood or fresh tissues. For formalin-fixed, paraffin-embedded (FFPE) tissues immunohistochemistry (IHC) can be used; however, it may not be as sensitive and serotyping is not possible. The application of reverse transcription-polymerase chain reaction (RT-PCR) for the detection of DENV in FFPE tissues has been very limited. We evaluated FFPE autopsy tissues of 122 patients with suspected DENV infection by flavivirus and DENV RT-PCR, sequencing, and DENV IHC. The DENV was detected in 61 (50%) cases by RT-PCR or IHC. The RT-PCR and sequencing detected DENV in 60 (49%) cases (DENV-1 in 16, DENV-2 in 27, DENV-3 in 8, and DENV-4 in 6 cases). No serotype could be identified in three cases. The IHC detected DENV antigens in 50 (40%) cases. The RT-PCR using FFPE tissue improves detection of DENV in fatal cases and provides sequence information useful for typing and epidemiologic studies.

Author Notes

*Address correspondence to Julu Bhatnagar, Infectious Diseases Pathology Branch, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (CDC), Bldg.18/SB-105, MS-G32, 1600 Clifton Road NE, Atlanta, GA 30333. E-mail: JBhatnagar@cdc.gov

Authors’ addresses: Julu Bhatnagar, Dianna M. Blau, Wun-Ju Shieh, Christopher D. Paddock, Clifton Drew, Lindy Liu, Tara Jones, Mitesh Patel, and Sherif R. Zaki, Infectious Diseases Pathology Branch, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (CDC), Atlanta, GA, E-mails: JBhatnagar@cdc.gov, DBlau@cdc.gov, WShieh@cdc.gov, CPaddock@cdc.gov, CDrew1@cdc.gov, LLiu1@cdc.gov, TJones5@cdc.gov, MPatel1@cdc.gov, and SZaki@cdc.gov.

Reprint requests: Julu Bhatnagar, Infectious Diseases Pathology Branch, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention (CDC), Bldg.18/SB-105, MS-G32, Atlanta, GA, Tel: 404-639-2826, Fax: 404-639-3043, E-mail: JBhatnagar@cdc.gov.

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