title: Development of a DNA microarray method for detection and identification of all 15 distinct O-antigen forms of Legionella pneumophila.
abstract: Legionella is ubiquitous in many environments. At least 50 species and 70 serogroups of the Gram-negative bacterium have been identified. Of the 50 species, 20 are pathogenic, and Legionella pneumophila is responsible for the great majority (approximately 90%) of the Legionnaires' disease cases that occur. Furthermore, of the 15 L. pneumophila serogroups identified, O1 alone causes more than 84% of the Legionnaires' disease cases that occur worldwide. Rapid and reliable assays for the detection and identification of L. pneumophila in water, environmental, and clinical samples are in great demand. L. pneumophila bacteria are traditionally identified by their O antigens by immunological methods. We have recently developed an O serogroup-specific DNA microarray for the detection of all 15 distinct O-antigen forms of L. pneumophila, including serogroups O1 to O15. A total of 35 strains were used to verify the specificity of the microarray, including 15 L. pneumophila O-antigen standard reference strains and seven L. pneumophila clinical isolates as target strains, seven reference strains of other non-pneumophila Legionella species as closely related strains, and six non-Legionella bacterial species as nonrelated strains. The detection sensitivity was 1 ng of genomic DNA or 0.4 CFU/ml in water samples with filter enrichment and plate culturing. This study demonstrated that the microarray allows specific, sensitive, and reproducible detection of L. pneumophila serogroups. To the best of our knowledge, this is the first report of a microarray serotyping method for all 15 distinct O-antigen forms of L. pneumophila.
authors: Key Laboratory of Molecular Microbiology and Technology of the Ministry of Education, TEDA College, Nankai University, TEDA, Tianjin, People's Republic of China.
authors: Boyang
authors: B
authors: Cao
authors: Fangfang
authors: F
authors: Yao
authors: Xiangqian
authors: X
authors: Liu
authors: Lu
authors: L
authors: Feng
authors: Lei
authors: L
authors: Wang
mesh_terms: D001483:Base Sequence; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D016952:Legionella pneumophila / Q000235:genetics*; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction; D019081:O Antigens / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010802:Phylogeny; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D019081:O Antigens
references: J Microbiol. 2012 Feb;50(1):127-36
references: 22367947
references: Int J Food Microbiol. 2011 Jan 31;145(1):293-300
references: 21276629
references: PLoS One. 2013;8(3):e59018
references: 23527075
references: Clin Infect Dis. 2002 Oct 15;35(8):990-8
references: 12355387
references: J Med Microbiol. 2009 Jan;58(Pt 1):69-81
references: 19074655
references: Arch Intern Med. 1997 Aug 11-25;157(15):1709-18
references: 9250232
references: J Microbiol Methods. 2008 Oct;75(2):329-34
references: 18700154
references: J Clin Microbiol. 2006 Oct;44(10):3608-15
references: 16943358
references: Clin Infect Dis. 2003 Jan 1;36(1):64-9
references: 12491204
references: Appl Microbiol Biotechnol. 2011 Aug;91(3):777-87
references: 21626022
references: Eur J Biochem. 1994 Apr 1;221(1):239-45
references: 8168511
references: Clin Microbiol Rev. 2002 Jul;15(3):506-26
references: 12097254
references: Vet Microbiol. 2005 Dec 20;111(3-4):181-90
references: 16280204
references: J Bacteriol. 1996 Sep;178(17):5205-14
references: 8752339
references: Appl Environ Microbiol. 2013 Jun;79(11):3392-9
references: 23524674
references: Curr Top Med Chem. 2008;8(2):141-51
references: 18289083
references: Appl Environ Microbiol. 2012 Jun;78(11):3966-74
references: 22447597
references: J Clin Microbiol. 2006 Dec;44(12):4376-83
references: 17021058
references: Appl Environ Microbiol. 2010 Dec;76(24):8201-10
references: 20971864
references: J Clin Microbiol. 2004 Jan;42(1):458-60
references: 14715805
references: N Engl J Med. 1977 Dec 1;297(22):1189-97
references: 335244
references: Int J Med Microbiol. 2000 Mar;290(1):37-49
references: 11043980
references: Nature. 2008 Nov 6;456(7218):53-9
references: 18987734
references: Appl Environ Microbiol. 2011 Apr;77(7):2209-14
references: 21296934
references: Infect Immun. 1998 Aug;66(8):3545-51
references: 9673232
references: J Clin Microbiol. 2009 May;47(5):1386-92
references: 19261788
references: J Bacteriol. 1996 Mar;178(6):1631-9
references: 8626291
references: FEMS Microbiol Rev. 2008 Jul;32(4):627-53
references: 18422615
references: Gene. 2012 Nov 1;509(1):142-8
references: 22967710
references: Curr Microbiol. 2008 Oct;57(4):294-300
references: 18587615
references: Mol Microbiol. 1994 Nov;14(3):505-19
references: 7533882