Simultaneous Detection of Pathogens in Clinical Samples from Patients with Community-Acquired Pneumonia by Real-Time PCR with Pathogen-Specific Molecular Beacon Probes

• DOI: 10.1128/jcm.44.4.1440-1446.2006
• Publication Date: 2006-04-06T13:42:51Z
• Authors: Miyuki Morozumi; Eiichi Nakayama; Satoshi Iwata; Yasuko Aoki; Keiko Hasegawa; Reiko Kobayashi; Naoko Chiba; Takeshi Tajima; Kimiko Ubukata
• Keywords: Chlamydophila pneumoniae; Molecular beacon

ABSTRACT In this study, real-time PCR with pathogen-specific molecular beacons (MB) and primers was evaluated for prediction of community-acquired pneumonia (CAP) causative agents, detecting six main CAP Streptococcus pneumoniae , Haemophilus influenzae Mycoplasma Chlamydophila Legionella pneumophila pyogenes simultaneously. The assay fresh clinical specimens from infants children ( n = 389) adults 40). MB probes are both pathogen specific, namely, the lytA gene S. mip L. 16S rRNA genes remaining four organisms. DNA extraction performed a commercially available EXTRAGEN II kit, amplification Stratagene Mx3000P. limit detection these pathogens ranged 2 copies to 18 copies. whole process analysis finished in less than h. obtained sensitivity specificity study relative those conventional cultures were as follows: 96.2% 93.2% 95.8% 95.4% H. 100% M. respectively. serologic 90.2% 97.9%, samples C. gave positive predictable values, cases, elevation titer value also observed. conclusion, we demonstrated that is useful identifying agents rapidly examining course empirical chemotherapy timely manner, supporting culture methods.