Korean J Clin Microbiol.  2010 Mar;13(1):40-46. 10.5145/KJCM.2010.13.1.40.

Clinical Evaluation of the Multiplex PCR Assay for the Detection of Bacterial Pathogens in Respiratory Specimens from Patients with Pneumonia

• Jung CL
• Lee MA
• Chung WS

• KMID: 1828594
• DOI: http://doi.org/10.5145/KJCM.2010.13.1.40

Abstract

BACKGROUND
Community-acquired pneumonia (CAP) is a major infectious disease with significant morbidity and mortality worldwide. Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis are common pathogens of CAP; however, the conventional methods used to detect these agents, including culturing, lack sensitivity and are time-consuming. We evaluated a recently developed multiplex PCR assay which can test these agents simultaneously.
METHODS
One hundred patients with pneumonia and 99 healthy adults were tested using the Seeplex Pneumobacter ACE Detection assay (Seegene, Inc., Seoul, Korea). Culture and urinary antigen tests were also performed.
RESULTS
In patients with pneumonia, the positive detection rates of PCR for S. pneumoniae and H. influenzae were 52.0% (52/100) and 30.0% (30/100), respectively, those of M. pneumoniae and L. pneumophila were 2.0% (2/100) and 1.0% (1/100), respectively, and B. pertussis and C. pneumoniae were not detected. In healthy adults, the detection rates of S. pneumoniae and H. influenzae revealed similar results, 53.5% (53/101) and 40.4% (40/101), respectively, and the other four pathogens were not detected. The sensitivity and specificity of PCR for S. pneumoniae in pneumonia patients were 100% (95% confidence interval [CI], 87.9~100%) and 65.7% (95% CI, 55.2~76.5%), respectively, according to the urinary antigen test and cultures of the respiratory samples and blood.
CONCLUSION
Differentiating S. pneumoniae and H. influenzae colonization from infection was difficult using the PCR assay. Therefore, the use of this assay is inappropriate for the diagnosis of pneumonia due to these agents, although multiplex PCR assay would be useful for the detection of M. pneumoniae and L. pneumophila.

Keyword

Multiplex PCR; Community-acquired pneumonia (CAP); Streptococcus pneumoniae; Haemophilus influenzae

MeSH Terms

Adult
Bordetella pertussis
Chlamydial Pneumonia
Chlamydophila pneumoniae
Colon
Communicable Diseases
Haemophilus influenzae
Humans
Influenza, Human
Legionella pneumophila
Multiplex Polymerase Chain Reaction
Mycoplasma pneumoniae
Pneumonia
Pneumonia, Mycoplasma
Polymerase Chain Reaction
Sensitivity and Specificity
Streptococcus pneumoniae
Whooping Cough

Figure

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  Fig. 1. The results for PCR products from the multiplex PCR on agarose gel. Lane 1, Molecular weight marker (350 bp band indicates Streptococcus pneumoniae and 257 bp band indicates Haemophilus influenzae); Lane 2, Negative; Lane 3 and 4, H. influenzae; Lane 5 and 6, S. pneumoniae and H. influenzae.

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