Detection of Brazilian hantavirus by reverse transcription polymerase chain reaction amplification of N gene in patients with hantavirus cardiopulmonary syndrome
Marcos Lázaro Moreli
Ricardo Luiz Moro de Sousa
Luiz Tadeu Moraes Figueiredo
- Publisher: Instituto Oswaldo Cruz, Ministério da Saúde
Memórias do Instituto Oswaldo Cruz.
(issn: 0074-0276, eissn: 1678-8060)
hantavirus cardiopulmonary syndrome | RC955-962 | Microbiology | nested RT-PCR | QR1-502 | N gene of hantavirus | Arctic medicine. Tropical medicine
mesheuropmc: viruses | virus diseases | animal diseases
We report a nested reverse transcription-polymerase chain reaction (RT-PCR) assay for hantavirus using primers selected to match high homology regions of hantavirus genomes detected from the whole blood of hantavirus cardiopulmonary syndrome (HCPS) patients from Brazil, also including the N gene nucleotide sequence of Araraquara virus. Hantavirus genomes were detected in eight out of nine blood samples from the HCPS patients by RT-PCR (88.9% positivity) and in all 9 blood samples (100% positivity) by nested-PCR. The eight amplicons obtained by RT-PCR (P1, P3-P9), including one obtained by nested-PCR (P-2) and not obtained by RT-PCR, were sequenced and showed high homology (94.8% to 99.1%) with the N gene of Araraquara hantavirus. Although the serologic method ELISA is the most appropriate test for HCPS diagnosis, the use of nested RT-PCR for hantavirus in Brazil would contribute to the diagnosis of acute hantavirus disease detecting viral genomes in patient specimens as well as initial genomic characterization of circulating hantaviruses.