Molecular detection and typing of dengue viruses from archived tissues of fatal cases by rt-PCR and sequencing: diagnostic and epidemiologic implications.
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.
Infectious Diseases Pathology Branch, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA. JBhatnagar@cdc.gov
Julu
J
Bhatnagar
Dianna M
DM
Blau
Wun-Ju
WJ
Shieh
Christopher D
CD
Paddock
Clifton
C
Drew
Lindy
L
Liu
Tara
T
Jones
Mitesh
M
Patel
Sherif R
SR
Zaki
D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D003715:Dengue / Q000175:diagnosis* / Q000453:epidemiology* / Q000821:virology; D003716:Dengue Virus / Q000145:classification* / Q000302:isolation & purification* / Q000472:pathogenicity; D005260:Female; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007223:Infant; D008297:Male; D008875:Middle Aged; D016612:Paraffin Embedding / Q000379:methods; D015995:Prevalence; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017423:Sequence Analysis, RNA; D012703:Serotyping; D013048:Specimen Handling; D016707:Tissue Fixation; D055815:Young Adult
D005404:Fixatives; D012367:RNA, Viral; D005557:Formaldehyde