Development and implementation of real-time nucleic acid amplification for the detection of enterovirus infections in comparison to rapid culture of various clinical specimens

Several real-time PCR and nucleic acid sequence-based amplification (NASBA) primer pairs and a modified real-time PCR primer pair for the detection of enteroviruses were compared. The modified real-time PCR primer pair was evaluated on clinical samples in comparison with cell culture using the Magna...

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Bibliographic Details
Published in:	Journal of medical virology Vol. 79; no. 12; pp. 1868 - 1876
Main Authors:	van Doornum, G.J.J, Schutten, M, Voermans, J, Guldemeester, G.J.J, Niesters, H.G.M
Format:	Journal Article
Language:	English
Published:	Hoboken Wiley Subscription Services, Inc., A Wiley Company 01-12-2007 Wiley-Liss
Subjects:	Biological and medical sciences Enterovirus Enterovirus - genetics Enterovirus - growth & development Enterovirus - isolation & purification Enterovirus Infections - cerebrospinal fluid Enterovirus Infections - diagnosis Enterovirus Infections - virology enteroviruses Feces - virology Fundamental and applied biological sciences. Psychology genotyping Human viral diseases Humans Infectious diseases Medical sciences Microbiology Miscellaneous Nucleic Acid Amplification Techniques - methods phenotyping real-time amplification Rhinovirus - genetics Rhinovirus - growth & development Rhinovirus - isolation & purification rhinoviruses Sensitivity and Specificity Viral diseases Virology Virus Cultivation - methods virus culture Typing Picornaviridae genotyping Genotype real-time amplification Enterovirus rhino- viruses Real time phenotyping virus culture Virus Amplification Infection enteroviruses Detection Clinical isolate Nucleic acid Comparative study
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Summary:	Several real-time PCR and nucleic acid sequence-based amplification (NASBA) primer pairs and a modified real-time PCR primer pair for the detection of enteroviruses were compared. The modified real-time PCR primer pair was evaluated on clinical samples in comparison with cell culture using the MagnaPure LC Isolation instrument for nucleic acid extraction. Six hundred forty samples could be examined both by cell culture and real-time PCR. Faecal specimens (n = 285), cerebrospinal fluid (n = 210), throat swabs (n = 113), biopsies (n = 1---, vesicular fluid (n = 11), and pleural fluid specimens (n = 9) were included. By culture, 26/640 (4%) samples were positive for enterovirus. By real-time PCR, the number of positive specimens was 50 (7.8%). Of the 210 cerebrospinal fluid samples, three were positive by culture and nine by real-time PCR. Seventeen and 33 of a total of 285 faecal specimens were positive by culture and real-time PCR, respectively. In case of discrepant results, the clinical symptoms were in accordance with an infection due to enteroviruses. Genotyping using the VP1 gene correlated with serotyping by neutralization. In contrast, six of the 19 specimens that could be typed both by neutralization and by sequencing using the VP4 domain yielded a different genotype, yet within the same species. Real-time PCR turned out to be suitable for the detection of enteroviruses in the daily routine setting. In comparison to rapid culture, it offers a rapid, more sensitive, and reliable assay; especially in cerebrospinal fluid, the yield of enteroviruses is much higher. J. Med. Virol. 79:1868-1876, 2007. © Wiley-Liss, Inc.
Bibliography:	http://dx.doi.org/10.1002/jmv.21031 ArticleID:JMV21031 istex:BA1A32F97A51315586C153BB96511C4C4D30E71F ark:/67375/WNG-3XW4RM45-R ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0146-6615 1096-9071
DOI:	10.1002/jmv.21031