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

Peptide Nucleic Acid–Based In Situ Hybridization Assay for Detection of Parvovirus B19 Nucleic... Abstract 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. © 2006 The American Association for Clinical Chemistry « Previous | Next Article » Table of Contents This Article Published March 30, 2006. doi: 10.1373/clinchem.2005.064741 Clinical Chemistry June 2006 vol. 52 no. 6 973-978 » Abstract Full Text PDF 064741.Supp Data Tables All Versions of this Article: clinchem.2005.064741v1 52/6/973 most recent Classifications Molecular Diagnostics and Genetics Services Email this article to a friend Alert me when this article is cited Alert me if a correction is posted Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Download to citation manager Responses No responses published Citing Articles Load citing article information Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Bonvicini, F. Articles by Gallinella, G. Search for related content PubMed PubMed citation Articles by Bonvicini, F. Articles by Gallinella, G. Related Collections Molecular Diagnostics and Genetics Related Content Load related web page information Follow Us Clinical Chemistry Trainee Council Register Today! www.traineecouncil.org Information for Authors Submit a Manuscript Editorial Board Clinical Case Studies Clinical Chemistry Guide to Scientific Writing Journal Club Podcasts Translated Content Annual Meeting Abstracts Permissions and Reprints Advertising Copyright © 2012 by the American Association for Clinical Chemistry http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Clinical Chemistry American Association for Clinical Chemistry

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

Loading next page...
 
/lp/american-association-for-clinical-chemistry/peptide-nucleic-acid-based-in-situ-hybridization-assay-for-detection-al07YRBEpB
Publisher
American Association for Clinical Chemistry
Copyright
Copyright © 2006 by the American Association for Clinical Chemistry.
ISSN
0009-9147
eISSN
1530-8561
D.O.I.
10.1373/clinchem.2005.064741
Publisher site
See Article on Publisher Site

Abstract

Abstract 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. © 2006 The American Association for Clinical Chemistry « Previous | Next Article » Table of Contents This Article Published March 30, 2006. doi: 10.1373/clinchem.2005.064741 Clinical Chemistry June 2006 vol. 52 no. 6 973-978 » Abstract Full Text PDF 064741.Supp Data Tables All Versions of this Article: clinchem.2005.064741v1 52/6/973 most recent Classifications Molecular Diagnostics and Genetics Services Email this article to a friend Alert me when this article is cited Alert me if a correction is posted Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Download to citation manager Responses No responses published Citing Articles Load citing article information Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Bonvicini, F. Articles by Gallinella, G. Search for related content PubMed PubMed citation Articles by Bonvicini, F. Articles by Gallinella, G. Related Collections Molecular Diagnostics and Genetics Related Content Load related web page information Follow Us Clinical Chemistry Trainee Council Register Today! www.traineecouncil.org Information for Authors Submit a Manuscript Editorial Board Clinical Case Studies Clinical Chemistry Guide to Scientific Writing Journal Club Podcasts Translated Content Annual Meeting Abstracts Permissions and Reprints Advertising Copyright © 2012 by the American Association for Clinical Chemistry

Journal

Clinical ChemistryAmerican Association for Clinical Chemistry

Published: Jun 1, 2006

There are no references for this article.

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off