Abstract
Background
We intended to evaluate the diagnostic usefulness of a multiplex reverse transcriptase-PCR (mRT-PCR) assay kit under dual priming oligonucleotide system (DPO) for the childhood acute respiratory tract infections.
Methods
Two hundred nasopharyngeal aspirates were taken from children ≤5 yr old admitted due to acute respiratory infections in 2004. Direct fluorescent antibody (FA) assays were performed with fresh specimens; then, mRT-PCRs for the detection of 12 respiratory viruses (Seeplex RV detection kit, SeeGene, Seoul, Korea) were tested with frozen specimens.
Results
FA assays for five common respiratory viruses showed positive results in 66 patients (33.0%), while mRT-PCR detected causative viruses in 112 patients (56.0%), including 16 co-infected cases (8.0%). A total of 129 viruses were identified: respiratory syncytial virus A/B (38.0%/7.8%), influenza virus A/B (10.1%/5.4%), parainfluenza virus 1/2/3 (7.0%/3.1%/7.8%), coronavirus 229E or NL63 (6.2%), human metapneumovirus (4.7%), adenovirus (4.7%), rhinovirus (3.9%), and coronavirus OC43 (1.6%).
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Fig. 1.
Two sets (A set in the left and B set in the right) of mRT-PCR using dual priming oligonucleotide systems in 6 clinical specimens. Lane MA, Size marker of A set; MB, Size marker of B; N, negative control; 1-6, Patients 1-6; Patient 1, Positive for RSV B; Patient 2, coronavirus 229E/NL63 and rhinovirus; Patient 3, influenza A; Patient 4, RSV B, Patient 5, negative; Patient 6, influenza A. Each lane presents internal control band (719 bp). Adv, Adenovirus (534 bp); MPV, Human metapneumovirus (469 bp); 229E, Corovirus 229E/NL63 (375 bp); PIV1, Parainfluenza virus 1 (324 bp); PIV2, Parainfluenza virus 2 (264 bp); PIV3, Parainfluenza virus 3 (219 bp); Flu A, Influenza A virus (516 bp); Flu B, Influenza B virus (455 bp); RSV B, Respiratory syncytial virus B (391 bp); Rhino, Human rhinovirus A (340 bp); RSV A, Respiratory syncytial virus A (273 bp); OC43, Coronavirus OC43 (231 bp).
![kjlm-27-420f1.tif](/upload/SynapseXML/0039kjlm/thumb/kjlm-27-420f1.gif)
Abbreviation: mRT-PCR, multiple reverse transcriptase PCR.
Fig. 2.
Monthly distribution of 12 respiratory viruses isolated from children with acute respiratory tract infections in 2004 (Co-infected cases were excluded).
![kjlm-27-420f2.tif](/upload/SynapseXML/0039kjlm/thumb/kjlm-27-420f2.gif)
Table 1.
Viruses identified in 200 nasopharyngeal aspirates obtained from children with acute respiratory tract infections by mRT-PCR and fluorescent antibody assays
Virus identified | N (%) of positive specimens, by the indicated method of detection (N=200) | Concordance of mRT-PCR and fluorescent Ab assay | ||||
---|---|---|---|---|---|---|
mRT-PCR | Fluorescent Ab assay | Both (+) | PCR (+) Ab (–) | PCR (–) Ab (+) | Concordance (%)∗ | |
RSV A | 49 (24.5) | 45 (22.5)† | 43 | 6 | 2 | 84.3 |
RSV B | 10 (5.0) | 2 (1.0)† | 2 | 8 | 0 | 20.0 |
Influenza virus A | 13 (6.5) | 6 (3.0) | 5 | 8 | 1 | 35.7 |
Influenza virus B | 7 (3.5) | 2 (1.0) | 0 | 7 | 2 | 0 |
Parainfluenza virus 1 | 9 (4.5) | 2 (1.0)† | 2 | 7 | 0 | 22.2 |
Parainfluenza virus 2 | 4 (2.0) | 0† | 0 | 4 | 0 | 0 |
Parainfluenza virus 3 | 10 (5.0) | 7 (3.5)† | 7 | 3 | 0 | 70.0 |
Adenovirus | 6 (3.0) | 2 (1.0) | 2 | 4 | 0 | 33.3 |
Coronavirus 229E/NL63 | 8 (4.0) | Not tested | ||||
Coronavirus OC43 | 2 (1.0) | Not tested | ||||
Human metapneumovirus | 6 (3.0) | Not tested | ||||
Rhinovirus | 5 (2.5) | Not tested | ||||
Total | 129 (56.0)† | 66 (33.0) | 61 | 47 | 5 | 54.0 |
∗ Corcordance rate (%)=(N of positive results by both of the two methods∗100)/(N of positive results by both of the two methods + N of positive results in multiplex RT-PCR only+N of positive results in fluorescent Ab assay only);
Table 2.
Disease entities in 200 children with acute respiratory tract infections and their causative agents detected by mRT-PCR∗