Comparison of sputum and nasopharyngeal swab specimens for molecular diagnosis of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila

Abstract

Background: Differentiation of atypical pathogens is important for community-acquired pneumonia (CAP). In this study, we compared sputum and nasopharyngeal swabs (NPS) for use in detection of Mycoplasma pneumoniae (MP), Chlamydophila pneumoniae (CP), and Legionella pneumophila (LP), using Seeplex PneumoBacter ACE Detection Assay (PneumoBacter; Seegene).

Methods: Sputum and NPS specimens were collected from patients in 15 hospitals. DNA was extracted from sputum using QIAamp DNA Stool Mini Kit (Qiagen) and from NPS using easyMAG (bioMérieux). Both types of specimens were evaluated by multiplex PCR using PneumoBacter. To determine the diagnostic performance of this assay, sputum samples were also tested using BD ProbeTec ET Atypical Pneumonia Assay (APA; Becton Dickinson).

Results: Among 217 sputum and NPS, 20 (9.2%), 2 (0.9%), and 0 sputum were positive for MP, LP, and CP, respectively, whereas 8 (3.7%) NPS were positive for MP. The sputum APA test yielded 186, 206, and 204 interpretable results for MP, LP, and CP, respectively. Of these, 21 (11.3%) were positive for MP, 2 (1.0%) were positive for LP, and 0 samples were positive for CP. Compared to APA, the sensitivity and specificity of the sputum assay for MP were 95.2% and 100.0%, respectively, whereas for the NPS assay, these were 38.1% and 93.9%. Sputum testing was more sensitive than NPS testing (P=0.002). For LP and CP diagnosis, PneumoBacter and APA tests agreed 100%.

Conclusions: Specimen type is crucial and sputum is preferred over NPS for simultaneous detection of MP, LP, and CP using multiplex PCR in CAP.

Keywords: Atypical pneumonia; Chlamydophila pneumoniae; Legionella pneumophila; Mycoplasma pneumoniae; Nasopharyngeal swab; Polymerase chain reaction; Sputum.

Fig. 1

Results of the multiplex PCR Seeplex PneumoBacter ACE Detection Assay. Lane M, amplicon size marker. Bands of 583 bp, 472 bp, and 146 bp are characteristic of Mycoplasma pneumoniae, Legionella pneumophila, and Chlamydophila pneumoniae, respectively. Lane 1, negative control; Lanes 2 and 3, nasopharyngeal swab (NPS) and sputum samples from an Mycoplasma pneumoniae-positive patient; Lanes 4 and 5, NPS and sputum samples from a Legionella pneumophila-positive patient; Lane 6, positive control.