title: Diagnosis of Upper and Lower Respiratory Tract Bacterial Infections with the Use of Multiplex PCR Assays.
abstract: The investigation of respiratory infections by molecular techniques provides important information about the epidemiology of respiratory disease, especially during the post-vaccination era. The objective of the present study was the detection of bacterial pathogens directly in clinical samples from patients with upper and lower respiratory tract infections using multiplex polymerase chain reaction (PCR) assays developed in our laboratory. Clinical samples taken over a three-year period (2007-2009) and obtained from 349 patients (adults (n = 66); children (n = 283)) with signs and symptoms of certain upper or lower respiratory tract infections, consisted of: bronchoalveolar lavages (BAL, n = 83), pleural fluids (n = 29), and middle-ear aspirates (n = 237). Overall, 212 samples (61%) were confirmed by culture and/or PCR. Among the positive samples, Streptococcus pneumoniae (mainly serotype 3) was predominant (104/212; 49.0%), followed by non-typable Haemophilus influenzae (NTHi) 59/212; 27.8%) and Streptococcus pyogenes (47/212; 22%). Haemophilus influenzae type b was detected in only three samples. The underlying microbiology of respiratory infections is gradually changing in response to various selective pressures, such as vaccine use and antibiotic consumption. The application of multiplex PCR (mPCR) assays is particularly useful since it successfully identified the microorganisms implicated in acute otitis media or lower respiratory tract infections in nearly 75% of patients with a positive result compared to conventional cultures. Non-culture identification of the implicated pneumococcal serotypes is also an important issue for monitoring pneumococcal infections in the era of conjugate pneumococcal vaccines.
authors.affiliation: National Meningitis Reference Laboratory, National School of Public Health, 196, Alexandras Avenue, Athens 11521, Greece. markoulatos@bio.uth.gr.|School of Health Sciences, Department of Biochemistry and Biotechnology, University of Thessaly, Ploutonos 26 & Aiolou str, Larissa 41221, Greece. markoulatos@bio.uth.gr.
authors.forename: Athanasia
authors.initials: A
authors.lastname: Xirogianni
authors.affiliation: Second Department of Pediatrics, University of Athens School of Medicine, P. and A. Kyriakou Children's Hospital, Thevon & Levadeias str, Athens 11527, Greece. matsolia@ath.forthnet.gr.
authors.forename: Maria
authors.initials: M
authors.lastname: Tsolia
authors.affiliation: Penteli's Children's Hospital, Microbiology Laboratory, 8 Ippokratous str., Penteli, Attiki 15236, Greece. grace.iliadou@gmail.com.
authors.forename: Aliki
authors.initials: A
authors.lastname: Voyiatzi
authors.affiliation: National Meningitis Reference Laboratory, National School of Public Health, 196, Alexandras Avenue, Athens 11521, Greece. msioumala78@yahoo.com.
authors.forename: Maria
authors.initials: M
authors.lastname: Sioumala
authors.affiliation: Penteli's Children's Hospital, Microbiology Laboratory, 8 Ippokratous str., Penteli, Attiki 15236, Greece. tmakri@gmail.com.
authors.forename: Antonia
authors.initials: A
authors.lastname: Makri
authors.affiliation: "Evaggelismos" General Hospital of Athens, Microbiology Laboratory, 45-47 Ipsilantou Str, Athens 10676, Greece. gatzea@otenet.gr.
authors.forename: Athina
authors.initials: A
authors.lastname: Argyropoulou
authors.affiliation: "Evaggelismos" General Hospital of Athens, Microbiology Laboratory, 45-47 Ipsilantou Str, Athens 10676, Greece. opaniara@otenet.gr.
authors.forename: Olga
authors.initials: O
authors.lastname: Paniara
authors.affiliation: School of Health Sciences, Department of Biochemistry and Biotechnology, University of Thessaly, Ploutonos 26 & Aiolou str, Larissa 41221, Greece. markoulatos@bio.uth.gr.
authors.forename: Panayotis
authors.initials: P
authors.lastname: Markoulatos
authors.affiliation: National Meningitis Reference Laboratory, National School of Public Health, 196, Alexandras Avenue, Athens 11521, Greece. kremastinou@esdy.edu.gr.
authors.forename: Jenny
authors.initials: J
authors.lastname: Kourea-Kremastinou
authors.affiliation: National Meningitis Reference Laboratory, National School of Public Health, 196, Alexandras Avenue, Athens 11521, Greece. gtzanakaki@esdy.edu.gr.
authors.forename: Georgina
authors.initials: G
authors.lastname: Tzanakaki
keywords: ear aspirates; molecular diagnosis; pleural fluids
references.citation: Diagn Microbiol Infect Dis. 2009 Feb;63(2):121-6
references.pmid: 19027257
references.citation: J Med Microbiol. 2005 Dec;54(Pt 12):1133-8
references.pmid: 16278425
references.citation: Emerg Infect Dis. 2008 Sep;14(9):1390-7
references.pmid: 18760005
references.citation: FEMS Immunol Med Microbiol. 2003 Oct 24;39(1):31-6
references.pmid: 14556993
references.citation: J Antimicrob Chemother. 2002 Dec;50 Suppl S2:59-73
references.pmid: 12556435
references.citation: J Infect Dis. 2004 Mar 1;189(5):775-84
references.pmid: 14976593
references.citation: Acta Otolaryngol Suppl. 1996;523:128-9
references.pmid: 9082757
references.citation: Arch Dis Child. 2008 Apr;93(4):316-8
references.pmid: 18006562
references.citation: J Clin Microbiol. 2002 Sep;40(9):3115-20
references.pmid: 12202540
references.citation: J Infect Chemother. 2008 Jun;14(3):180-7
references.pmid: 18574652
references.citation: Eur Respir J. 2001 Apr;17(4):791-801
references.pmid: 11401077
references.citation: Clin Microbiol Infect. 2005 May;11(5):386-90
references.pmid: 15819865
references.citation: J Clin Microbiol. 1998 May;36(5):1185-8
references.pmid: 9574673
references.citation: J Infect Dis. 2010 Jan 1;201(1):32-41
references.pmid: 19947881
references.citation: Eur Respir J. 2006 Sep;28(3):568-75
references.pmid: 16737990
references.citation: Pediatrics. 2010 Jan;125(1):26-33
references.pmid: 19948570
references.citation: J Clin Microbiol. 2003 Jun;41(6):2378-84
references.pmid: 12791852
references.citation: Vaccine. 2008 Dec 23;26 Suppl 7:G5-10
references.pmid: 19094935
references.citation: Curr Opin Pulm Med. 2010 May;16(3):217-25
references.pmid: 20375783
references.citation: JAMA. 1995 May 24-31;273(20):1598-604
references.pmid: 7745773
references.citation: Paediatr Respir Rev. 2007 Sep;8(3):204-10, quiz 210-1
references.pmid: 17868918
references.citation: J Clin Microbiol. 1998 Dec;36(12):3605-8
references.pmid: 9817881
references.citation: J Infect Dis. 1997 Jun;175(6):1440-5
references.pmid: 9180184
references.citation: J Clin Microbiol. 1995 Jul;33(7):1835-8
references.pmid: 7665655
references.citation: APMIS. 2005 Feb;113(2):99-111
references.pmid: 15723684
references.citation: Clin Infect Dis. 2005 Jul 1;41(1):35-41
references.pmid: 15937760
references.citation: Res Microbiol. 2005 May;156(4):564-7
references.pmid: 15862455