Journal List > Korean J Clin Microbiol > v.12(3) > 1038162

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

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.

Conclusion

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.

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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.
kjcm-12-110f1.tif
Table 1.
Detection of 7 major respiratory viruses in nasopharyngeal aspirates (n=122) by DFA and mRT-PCR methods
DFA mRT-PCR
ADV 11 20 (PIV3)
FluA 10 12 (ADV)
FluB 1 4
PIV1 5 8 (PIV3)
PIV2 0 1
PIV3 9 17 (PIV2)
RSV 3 (ADV) 6 (ADV)
Negative 83 54

Include 1 case of mixed infection with the virus within the parenthesis.

Abbreviations: ADV, adenovirus; Flu, influenza virus A, B; PIV, parainfluenza virus 1, 2, 3; RSV, respiratory syncytial virus.

Table 2.
Detection of 7 major respiratory viruses in nasopharyngeal apirates (n=85) by shell vial culture (SVC) and direct immunofluorescence assay (DFA)
DFA SVC
ADV
FluA
PIV1
PIV3
RSV
6 (RSV)
2
2
6
12
16
1
3 (ADV)
6 (ADV)
8
Negative 57 51

Include 1 case of mixed infection with the virus within the parenthesis.

Abbreviations: See Table 1.

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