Tf. Schwarz et al., DETECTION AND SEQUENCE CONFIRMATION OF SIN-NOMBRE VIRUS-RNA IN PARAFFIN-EMBEDDED HUMAN TISSUES USING ONE-STEP RT-PCR, Journal of virological methods, 51(2-3), 1995, pp. 349-356
Citations number
22
Categorie Soggetti
Virology,"Biochemical Research Methods","Biothechnology & Applied Migrobiology
Sin Nombre virus (SNV) is the causative agent of hantavirus pulmonary
syndrome (HPS). SNV RNA can be detected in fresh or frozen autopsy tis
sue samples by reverse transcriptase polymerase chain reaction (RT-PCR
), and virus antigens can be identified by immunohistochemistry. A met
hod was developed for demonstration of SNV RNA in formalin-fixed, para
ffin-embedded tissues by RT-PCR. Virus genomes were detected in 8 of 1
2 (66.7%) fixed human tissue samples that were positive by immunohisto
chemistry, and RT-PCR prior to tissue fixation. By comparison with nuc
leotide sequences determined previously in fresh or frozen tissues of
the same patients, identical genomic sequences were obtained, proving
the authenticity of the PCR products. The study demonstrates that dete
ction of SNV RNA in formalin-fixed, paraffin-embedded archival tissue
by RT-PCR is feasible. This method allows retrospective studies on the
phylogenetic epidemiology of SNV in North America.