Utilization of autopsy RNA for the synthesis of the nucleocapsid antigen of a newly recognized virus associated with hantavirus pulmonary syndrome

Heinz Feldmann, Anthony Sanchez, Sergey Morzunov, Christina F. Spiropoulou, Pierre E. Rollin, Thomas Ksiazek, Clarence J. Peters, Stuart T. Nichol

Research output: Contribution to journalArticle

171 Citations (Scopus)

Abstract

A newly recognized hantavirus was recently found to be associated with an outbreak of acute respiratory illness in the southwestern United States. The disease, which has become known as hantavirus pulmonary syndrome, has an unusually high mortality (64%). Virus isolation attempts have been unsuccessful thus far, resulting in a lack of homologous antigen for use in diagnostic assays. For this reason, a molecular approach was initiated to produce recombinant homologous antigen. The virus nucleocapsid (N) protein was selected, since N has been shown to be a sensitive antigenic target in other hantavirus systems. The N protein open reading frame of the virus S genome segment was synthesized from frozen autopsy tissue by polymerase chain reaction amplification, followed by cloning and expression in Hela cells (vaccinia-T7 RNA polymerase system) and Escherichia coli. N protein-expressing Hela cells served as excellent antigens for an improved indirect immunofluorescence assay. Use of the E.coli-expressed N protein in an enzyme-linked immunosorbent assay improved the sensitivity and specificity when compared with heterologous antigens used previously. Preliminary analysis also indicates that the higher sensitivity could result in earlier detection of infected persons. These data demonstrate that even in the absence of a virus isolate, the necessary homologous antigen can be produced and can serve to improve the detection and diagnostic capabilities needed to combat this newly recognized fatal respiratory illness in the United States.

Original languageEnglish (US)
Pages (from-to)351-367
Number of pages17
JournalVirus Research
Volume30
Issue number3
DOIs
StatePublished - 1993
Externally publishedYes

Fingerprint

Hantavirus Pulmonary Syndrome
Nucleocapsid
Autopsy
RNA
Hantavirus
Viruses
Antigens
HeLa Cells
Southwestern United States
Heterophile Antigens
Escherichia coli
Nucleocapsid Proteins
Vaccinia
Proteins
Indirect Fluorescent Antibody Technique
Open Reading Frames
Disease Outbreaks
Organism Cloning
Enzyme-Linked Immunosorbent Assay
Genome

Keywords

  • Antigen
  • Hantavirus
  • Hantavirus pulmonary syndrome
  • Nucleocapsid protein
  • Recombinant expressed N protein

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

Cite this

Utilization of autopsy RNA for the synthesis of the nucleocapsid antigen of a newly recognized virus associated with hantavirus pulmonary syndrome. / Feldmann, Heinz; Sanchez, Anthony; Morzunov, Sergey; Spiropoulou, Christina F.; Rollin, Pierre E.; Ksiazek, Thomas; Peters, Clarence J.; Nichol, Stuart T.

In: Virus Research, Vol. 30, No. 3, 1993, p. 351-367.

Research output: Contribution to journalArticle

Feldmann, Heinz ; Sanchez, Anthony ; Morzunov, Sergey ; Spiropoulou, Christina F. ; Rollin, Pierre E. ; Ksiazek, Thomas ; Peters, Clarence J. ; Nichol, Stuart T. / Utilization of autopsy RNA for the synthesis of the nucleocapsid antigen of a newly recognized virus associated with hantavirus pulmonary syndrome. In: Virus Research. 1993 ; Vol. 30, No. 3. pp. 351-367.
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