Korean J Clin Microbiol.  2009 Sep;12(3):110-115. 10.5145/KJCM.2009.12.3.110.

Detection of Respiratory Viruses in Children by Multiplex Reverse Transcriptase PCR, Direct Immunofluorescence Assay, and Shell Vial Culture

• Yoon KH
• Cho JH

• KMID: 1828580
• DOI: http://doi.org/10.5145/KJCM.2009.12.3.110

Abstract

BACKGROUND
Direct immunofluorescence assay (DFA) and shell vial culture (SVC) have been used to diagnose respiratory viral infections. Recently a multiplex reverse transcriptase PCR (mRT-PCR) for 12 respiratory viruses has been introduced. We evaluated the diagnostic usefulness of these methods.
METHODS
Among 275 nasopharyngeal aspirates (NPAs) received from pediatric patients during the 3-month period from May through July, 2007, 122 samples were selected so as to include diverse viruses and varying numbers of DFA-positive cells for mRT-PCR. Also, the results of the 85 NPAs that had been analyzed by both DFA and SVC were reviewed retrospectively.
RESULTS
Detection rates for the seven major respiratory viruses, respiratory syncytial virus (RSV), influenza virus A and B, parainfluenza virus 1, 2, and 3, and adenovirus by DFA vs mRT-PCR were 32.0% and 55.7%, and by DFA vs SVC were 32.9% and 40.0%. A number of adenovirus detected by DFA vs mRT-PCR were 12 and 22, and by DFA vs SVC were 6 and 18. A number of RSV detected were 3 and 6, and 13 and 8, respectively.
CONCLUSIONS
mRT-PCR detected the respiratory viruses at the highest rate, followed by SVC and DFA in a decreasing order. However, DFA and multiplex PCR were more sensitive than SVC for RSV, while SVC was more sensitive than the other methods for adenovirus.

Keyword

Direct immunofluorescence assay; Shell vial culture; Multiplex PCR; Respiratroy virus

MeSH Terms

Adenoviridae
Child
Fluorescent Antibody Technique
Fluorescent Antibody Technique, Direct
Humans
Multiplex Polymerase Chain Reaction
Orthomyxoviridae
Paramyxoviridae Infections
Respiratory Syncytial Viruses
Reverse Transcriptase Polymerase Chain Reaction
RNA-Directed DNA Polymerase
Viruses
RNA-Directed DNA Polymerase

Figure

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  Fig. 1. Electrophoresis of mRT-PCR products for 12 repiratory viruses. Abbreviations: Control, internal control; ADV, adenovirus; HMPV, human metapneumo-virus; 229E, coronavirus 229E/NL63; PIV1, PIV2, PIV3, parainfluenza virus 1, 2, 3; FluA, FluB, influenza virus A, B; RSVA, B, respiratory syncytial virus A, B; HRVA, human rhinovirus A; OC43, coronavirus OC43. M1, 100 bp size marker; M2, viruses marker; NC, negative control; lane 1∼13, patients: patient 1 HRVA, patient 2 HRVA, patient 3 ADV, patient 5 PIV1, patient 7 PIV3, patient 9 RSVA, patient 10 PIV1, patient 12 ADV+RSVA, and patient 13 PIV1. Other patients were negative.

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