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Abstract
Background:
Rapid identification of the causative agent among potential bacterial and viral pathogens is important for the management of acute respiratory disease. In this study, we evaluated the analytical performance and clinical usefulness of a recently-introduced multiplex PCR assay, Seeplex™ Pneumobacter detection kit (Seegene Inc., Korea) for the identification of respiratory bacterial pathogens.
Methods:
One hundred and eighty one nasopharyngeal aspirates were collected from pediatric patients with respiratory symptoms and analysed by multiplex PCR for the detection of Streptococcus pneumoniae (S.P), Haemophilus influenzae (H.I), Mycoplasma pneumoniae (M.P), Chlamydophila pneumoniae (C.P), Bordetella pertussis (B.P) and Legionella pneumophila (L.P). A comparison of multiplex PCR with conventional culture for the isolation of S.P and H.I was performed on 112 specimens. The cross reactivity of multiplex PCR was also evaluated.
Results:
Of 181 cases, 81 cases were positive by multiplex PCR (44.8%): 52 cases for S.P (28.7%), 47 cases for H.I (26.0%), 9 cases for M.P (5.0%), 3 cases for B.P (1.7%) and 1 case for C.P (0.6%) including multiple infection cases. The agreement rates between multiplex PCR and culture for S.P and H.I were 92.9% (kappa index=0.84, P<0.001) and 91.1% (kappa index=0.75, P<0.001), respectively. There was no cross reactivity with common bacterial and viral pathogens.
Conclusions:
Seeplex™ Pneumobacter detection kit could be a useful screening tool for the rapid detection of respiratory bacterial pathogens. Further studies with lower respiratory tract specimens would be needed for the clinical evaluation of S. pneumoniae and H. influenzae detected by multiplex PCR.
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Fig. 1.
Multiplex PCR products of six positive samples. M, 100 bp DNA ladder; lane 1, M. pneumoniae (583 bp); lane 2, L. pneumophila (472 bp); lane 3, S. pneumoniae (350 bp); lane 4, H. influenzae (257 bp); lane 5, B. pertussis (200 bp), lane 6, C. pneumoniae (146 bp); P, positive control ladder; N, negative control.
Table 1.
Target genes used in multiplex PCR
Table 2.
Results of analysis on 181 nasopharyngeal aspirates by multiplex PCR
Table 3.
Analytical specificity of multiplex PCR with 21 different microorganisms
Table 4.
Results of multiplex PCR for potential respiratory bacterial and viral pathogens
Table 5.
Comparison of multiplex PCR and conventional culture method for the detection of S. pneumoniae and H. influenzae in 112 patients
Table 6.
Disease entities of positive cases by multiplex PCR
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