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Development and implementation of real-time nucleic acid amplification for the detection of enterovirus infections in comparison to rapid culture of various clinical specimens.
- Source :
-
Journal of medical virology [J Med Virol] 2007 Dec; Vol. 79 (12), pp. 1868-76. - Publication Year :
- 2007
-
Abstract
- 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.<br /> ((c) Wiley-Liss, Inc.)
- Subjects :
- Enterovirus genetics
Enterovirus growth & development
Enterovirus Infections cerebrospinal fluid
Feces virology
Humans
Rhinovirus genetics
Rhinovirus growth & development
Rhinovirus isolation & purification
Sensitivity and Specificity
Enterovirus isolation & purification
Enterovirus Infections diagnosis
Enterovirus Infections virology
Nucleic Acid Amplification Techniques methods
Virus Cultivation methods
Subjects
Details
- Language :
- English
- ISSN :
- 0146-6615
- Volume :
- 79
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Journal of medical virology
- Publication Type :
- Academic Journal
- Accession number :
- 17935176
- Full Text :
- https://doi.org/10.1002/jmv.21031