title: The use of multiplex real-time PCR improves the detection of the bacterial etiology of community acquired pneumonia.
abstract: Community-acquired pneumonia (CAP) is still a major cause of morbidity and   mortality especially to children and compromised hosts, such as the old and those with   underlying chronic diseases. Knowledge of pathogens causing CAP constitutes the basis for   selection of antimicrobial treatment. Previous data have shown that etiological agents can   be identified in only up to 50% of patients, but this figure can be improved by using polymerase   chain reaction (PCR). This study was designed to evaluate multiplex real-time PCR as a   method for rapid differential detection of five bacterial causes of CAP (Streptococcus   pneumoniae, Burkholderia pseudomallei and atypical bacterial pathogens namely Mycoplasma   pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila) in CAP patients   attending Hospital Tengku Ampuan Afzan (HTAA)/ Kuantan, Pahang, Malaysia. Two previously   developed multiplex real-time PCR assays, duplex for the differential detection of S.   pneumoniae and B. pseudomallei and triplex for the atypical bacterial pathogens, were used   to detect a bacterial cause of CAP in blood and respiratory samples. Thus, 46 blood and 45   respiratory samples collected from 46 adult CAP patients admitted to HTAA were analysed by   multiplex real-time PCR assays and conventional methods. The microbial etiology of CAP   could be established for 39.1% (18/46) of CAP patients by conventional methods and this was   increased to 65.2% (30/46) with the additional use of real-time PCR. The most frequently   detected pathogens were S. pneumoniae (21.7% - all by PCR alone), Klebsiella pneumoniae   (17.3%), B. pseudomallei (13% - 83% of them positive by PCR alone and 17% by both culture   and PCR), Pseudomonas aeruginosa (6.5%), M. pneumoniae (6.5% - all by serology), C.   pneumoniae (4.3% - all positive by both PCR and serology), L. pneumophila (2.1% - all by PCR   alone), Escherichia coli (4.3%). Haemophilus infuenzae, Acinetobacter lwoffii and   Acinetobacter baumannii were detected by conventional methods (2.1% for each).
authors.affiliation: Department of Basic Medical Sciences, Kulliyyah of Medicine, International Islamic University Malaysia. imad@iium.edu.my
authors.forename: M I A
authors.initials: MI
authors.lastname: Mustafa
authors.forename: F
authors.initials: F
authors.lastname: Al-Marzooq
authors.forename: S H
authors.initials: SH
authors.lastname: How
authors.forename: Y C
authors.initials: YC
authors.lastname: Kuan
authors.forename: T H
authors.initials: TH
authors.lastname: Ng
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001431:Bacteriological Techniques / Q000379:methods*; D001769:Blood / Q000382:microbiology; D017714:Community-Acquired Infections / Q000175:diagnosis*; D005260:Female; D006801:Humans; D008296:Malaysia; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D018410:Pneumonia, Bacterial / Q000175:diagnosis*; D015995:Prevalence; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D055815:Young Adult