The rtxA toxin gene of Kingella kingae: a pertinent target for molecular diagnosis of osteoarticular infections.
Kingella kingae is an emerging osteoarticular pathogen in young children. Its isolation by traditional culture methods remains difficult, underscoring the need to implement other diagnostic methods for its detection and identification, such as nucleic acid amplification tests. Although the genome of this bacterium has not yet been sequenced, a toxin named RTX has been identified. The goal of this study was to develop sensitive, specific, and rapid molecular methods based on the rtxA toxin gene sequence to diagnose this infection. Two real-time PCR assays (SYBR green and TaqMan chemistries) targeting this gene are reported. Sensitivity and specificity were first evaluated successfully with 67 strains: 31 Kingella kingae isolates and 36 strains from other bacterial species. Then, 52 clinical specimens positive or negative by culture and/or PCR (16S rRNA and cpn60 genes) were tested with these assays. A nested PCR assay with subsequent sequencing was also developed to confirm the presence of Kingella kingae isolates in these clinical specimens. The results obtained demonstrate that these assays are accurate for the diagnosis of Kingella kingae infection.
CHU de Bordeaux, Hopital Pellegrin, Laboratoire de Bactériologie, Bordeaux, France.
Philippe
P
Lehours
Anne-Marie
AM
Freydière
Olivier
O
Richer
Christophe
C
Burucoa
Sandrine
S
Boisset
Philippe
P
Lanotte
Marie Françoise
MF
Prère
Agnès
A
Ferroni
Christine
C
Lafuente
Francois
F
Vandenesch
Francis
F
Mégraud
Armelle
A
Ménard
D001427:Bacterial Toxins / Q000235:genetics*; D001431:Bacteriological Techniques / Q000379:methods*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D017882:Kingella kingae / Q000235:genetics*; D008969:Molecular Sequence Data; D016870:Neisseriaceae Infections / Q000175:diagnosis* / Q000382:microbiology; D010003:Osteoarthritis / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D001427:Bacterial Toxins; D004269:DNA, Bacterial
Nucleic Acids Res. 1988 Nov 25;16(22):10881-90
2849754
Clin Microbiol Infect. 2007 Apr;13(4):419-23
17359327
J Pediatr Orthop. 2010 Apr-May;30(3):301-4
20357599
J Clin Microbiol. 2010 Aug;48(8):3021-3
20573877
Curr Opin Rheumatol. 2008 Jul;20(4):463-70
18525362
Ann Biol Clin (Paris). 2008 Jul-Aug;66(4):454-8
18725349
Pediatr Infect Dis J. 2010 Jul;29(7):639-43
20182400
Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
9254694
Pediatr Infect Dis J. 2005 Aug;24(8):692-6
16094222
Pediatr Infect Dis J. 2007 May;26(5):377-81
17468645
J Clin Microbiol. 2009 Jun;47(6):1837-41
19369442
Infect Immun. 1992 Jul;60(7):2976-85
1612762
Arch Pediatr. 2007 Oct;14 Suppl 2:S91-6
17956825
J Bacteriol. 2007 Jan;189(2):430-6
17098895
Lancet Infect Dis. 2004 Jun;4(6):358-67
15172344
Emerg Infect Dis. 2000 Jan-Feb;6(1):85-7
10653579
Pediatr Infect Dis J. 2003 Sep;22(9):837-9
14515832
J Microbiol Methods. 2007 Jan;68(1):88-93
16904782
J Clin Microbiol. 1992 May;30(5):1278-81
1583131
Clin Infect Dis. 2008 May 15;46(10):1631-2
18419507
Clin Infect Dis. 1998 Nov;27(5):1328-9
9827297
Clin Infect Dis. 1997 May;24(5):860-6
9142783
Arch Pediatr. 2000 Sep;7(9):927-32
11028199
J Gen Microbiol. 1968 May;51(3):377-85
5185818
J Med Microbiol. 2009 Jan;58(Pt 1):65-68
19074654