Comparison of Sputum and Nasopharyngeal Swab Specimens for Molecular Diagnosis of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila

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 (bioMerieux). 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.

Serial Testing of T-SPOT.TB Assays with Anti-Tuberculosis Therapy in Patients with Extrapulmonay Tuberculosis / 감염과화학요법

BACKGROUND: Limited data are available for the clinical utility of serial interferon-gamma producing T-cell response after initiation of treatment in patients with extrapulmonary tuberculosis (TB). We studied the serial TB-specific antigen T-cell responses measured using the T-SPOT.TB assay during the course of therapy. MATERIALS AND METHODS: We prospectively enrolled adult patients who were newly diagnosed with active extrapulmonary TB over a 24-month period. All patients were given standard anti-TB treatment. Blood samples were obtained for T-SPOT.TB at diagnosis, as well as 1-, 3-, 6-, and 12-months after initiating anti-TB therapy. RESULTS: A total of 52 patients with extrapulmonary TB (38 confirmed and 14 probable TB) were included in the final analysis. All patients had clinical and radiologic improvement after treatment and cured. T-SPOT.TB was positive for 90% at diagnosis, 100% at 1-, 3-, and 6-months, and 93% at 12-months after initiation of anti-TB therapy. There was no significant difference in median T-cell response between early secreting antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) at all time points. Median T-cell response steadily increased up to 6 months and then decreased. CONCLUSIONS: T-SPOT.TB assay remained positive after successful anti-TB treatment in most patients with extrapulmonary TB. Our data suggests that serial T-SPOT.TB has limited clinical utility as a surrogate marker of treatment response in patients with extrapulmonary TB.