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Clinical Chemistry 52: 973-978, 2006. First published March 30, 2006; 10.1373/clinchem.2005.064741
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(Clinical Chemistry. 2006;52:973-978.)
© 2006 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Peptide Nucleic Acid–Based In Situ Hybridization Assay for Detection of Parvovirus B19 Nucleic Acids

Francesca Bonvicini, Claudia Filippone, Elisabetta Manaresi, Giovanna Angela Gentilomi, Marialuisa Zerbini, Monica Musiani and Giorgio Gallinellaa

1 Department of Clinical and Experimental Medicine, Division of Microbiology, University of Bologna, Bologna, Italy.

aAddress correspondence to this author at: Department of Clinical and Experimental Medicine, Division of Microbiology, University of Bologna, Via Massarenti 9, 40138 Bologna, Italy. Fax 39-51-307397; e-mail giorgio.gallinella{at}unibo.it.

Background: Peptide nucleic acid (PNA) molecules are known to bind complementary nucleic acid sequences with a much stronger affinity and with more stable binding than DNA or RNA molecules. We chose parvovirus B19, which is diagnosed by detection of nucleic acids by in situ hybridization assay (ISH) and/or PCR, as an experimental model to develop an ISH assay that uses biotinylated PNA probes to detect viral genome in clinical specimens.

Methods: We first optimized the PNA-ISH assay on B19-infected and mock-infected UT-7/EpoS1 cells and then tested the assay on archival B19 specimens and on consecutive specimens. All data were compared with data obtained with a standardized DNA-based ISH assay and confirmed by a PCR-ELISA.

Results: PNA-ISH detected B19 genome in a higher number of B19-infected UT-7/EpoS1 cells and with a more defined localization of viral nucleic acids than the standardized DNA-ISH assay. Moreover, PNA-ISH was able to detect B19 genome in all positive archival samples, whereas DNA-ISH failed in 5 samples. PNA-ISH detected more positive samples than DNA-ISH when consecutive specimens were analyzed, and a close agreement was found with PCR-ELISA results.

Conclusions: The PNA-ISH assay had sensitivity and specificity comparable to a PCR assay and was more practical and quicker to perform than standard hybridization assays. The assay may be a suitable diagnostic test for the detection of viral nucleic acids in clinical specimens.




The following articles in journals at HighWire Press have cited this article:


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 Nucleic Acids ResHome page
A. M. Blanco, L. Rausell, B. Aguado, M. Perez-Alonso, and R. Artero
A FRET-based assay for characterization of alternative splicing events using peptide nucleic acid fluorescence in situ hybridization
Nucleic Acids Res., September 1, 2009; 37(17): e116 - e116.
[Abstract] [Full Text] [PDF]

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 J. Clin. Microbiol.Home page
L. Pironi, F. Bonvicini, P. Gionchetti, A. D'Errico, F. Rizzello, C. Corsini, L. Foroni, and G. Gallinella
Parvovirus B19 Infection Localized in the Intestinal Mucosa and Associated with Severe Inflammatory Bowel Disease
J. Clin. Microbiol., May 1, 2009; 47(5): 1591 - 1595.
[Abstract] [Full Text] [PDF]


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