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Journal of Clinical Microbiology, January 2006, p. 166-171, Vol. 44, No. 1
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.1.166-171.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Detection of Rhinoviruses by Tissue Culture and Two Independent Amplification Techniques, Nucleic Acid Sequence-Based Amplification and Reverse Transcription-PCR, in Children with Acute Respiratory Infections during a Winter Season

K. Loens,1* H. Goossens,1 C. de Laat,2 H. Foolen,2 P. Oudshoorn,2 S. Pattyn,1 P. Sillekens,2 and M. Ieven1

Department of Medical Microbiology, University of Antwerp, Antwerp, Belgium,1 bioMérieux, Boxtel, The Netherlands2

Received 18 July 2005/ Returned for modification 15 September 2005/ Accepted 12 October 2005

Five hundred seventeen consecutive nasopharyngeal aspirates were collected between October 1998 and May 1999 for episodes of acute respiratory tract infections in children presenting at the University Hospital of Antwerp. Culture and nucleic acid amplification techniques—nucleic acid sequence-based amplification (NASBA) and reverse transcription-PCR (RT-PCR)—were applied to detect rhinoviruses (RVs). Other respiratory viruses were detected by immunofluorescence (IF) analysis of the specimens and IF analysis of shell vial cultures. Among the 517 specimens, 219 viral agents were identified. They were, in decreasing order, rhinoviruses (93 [18.0%]), respiratory syncytial virus (76 [14.7%]), adenoviruses (16 [3.1%]), influenza viruses (15 [2.9%]), enteroviruses (15 [2.9%]), and herpes simplex virus (4 [0.8%]). For the evaluation of rhinovirus detection, culture positivity and/or a positive reaction in the two independent amplification methods was used as an expanded "gold standard." Based on this standard, the sensitivity, specificity, positive predictive value, and negative predictive value of culture were 44.7, 100, 100, and 99.8%, and those of NASBA and RT-PCR were 85.1, 98.3, 83.3, and 98.5% and 82.9, 93.4, 55.7, and 98.2%, respectively. NASBA and RT-PCR produced comparable results and were significantly more sensitive than virus culture. RVs showed the highest incidence in acute respiratory tract infections in children.


* Corresponding author. Mailing address: Department of Medical Microbiology, University of Antwerp, Universiteitsplein 1 S3, B-2610 Wilrijk, Belgium. Phone: 32-3-820-25-51. Fax: 32-3-820-26-63. E-mail:katherine.loens{at}ua.ac.be.


Journal of Clinical Microbiology, January 2006, p. 166-171, Vol. 44, No. 1
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.1.166-171.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.




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