Distribution of the leptospiral immunoglobulin-like (lig) genes in pathogenic Leptospira species and application of ligB to typing leptospiral isolates
The family of leptospiral immunoglobulin-like (lig) genes comprises ligA, ligB and ligC. This study used PCR to demonstrate the presence of lig genes among serovars from a collection of leptospiral strains and clinical isolates. Whilst ligA and ligC appeared to be present in a limited number of pathogenic serovars, the ligB gene was distributed ubiquitously among all pathogenic strains. None of the lig genes were detected among intermediate or saprophytic Leptospira species. It was also shown that, similar to the previously characterized secY gene, a short specific PCR fragment of ligB could be used to correctly identify pathogenic Leptospira species. These findings demonstrate that ligB is widely present among pathogenic strains and may be useful for their reliable identification and classification.
AhmedN.,
DeviS. M.,
ValverdeM. A.,
VijayachariP.,
Machang'uR. S.,
EllisW. A.,
HartskeerlR. A.2006; Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species. Ann Clin Microbiol Antimicrob 5:28[CrossRef]
BarocchiM. A.,
KoA. I.,
FerrerS. R.,
FariaM. T.,
ReisM. G.,
RileyL. W.2001; Identification of new repetitive element in Leptospira interrogans serovar Copenhageni and its application to PCR-based differentiation of Leptospira serogroups. J Clin Microbiol 39:191–195[CrossRef]
BhartiA. R.,
NallyJ. E.,
RicaldiJ. N.,
MatthiasM. A.,
DiazM. M.,
LovettM. A.,
LevettP. N.,
GilmanR. H.,
WilligM. R. & other authors; 2003; Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 3:757–771[CrossRef]
BrennerD. J.,
KaufmannA. F.,
SulzerK. R.,
SteigerwaltA. G.,
RogersF. C.,
WeyantR. S.1999; Further determination of DNA relatedness between serogroups and serovars in the family Leptospiraceae with a proposal for Leptospira alexanderi sp. nov. and four new Leptospira genomospecies. Int J Syst Bacteriol 49:839–858[CrossRef]
BrownP. D.,
LevettP. N.1997; Differentiation of Leptospira species and serovars by PCR-restriction endonuclease analysis, arbitrarily primed PCR and low-stringency PCR. J Med Microbiol 46:173–181[CrossRef]
BulachD. M.,
ZuernerR. L.,
WilsonP.,
SeemannT.,
McGrathA.,
CullenP. A.,
DavisJ.,
JohnsonM.,
KuczekE.other authors2006; Genome reduction in Leptospira borgpetersenii reflects limited transmission potential. Proc Natl Acad Sci U S A 103:14560–14565[CrossRef]
ChoyH. A.,
KelleyM. M.,
ChenT. L.,
MollerA. K.,
MatsunagaJ.,
HaakeD. A.2007; Physiological osmotic induction of Leptospira interrogans adhesion: LigA and LigB bind extracellular matrix proteins and fibrinogen. Infect Immun 75:2441–2450[CrossRef]
CrodaJ.,
FigueiraC. P.,
WunderE. A.Jr,
SantosC. S.,
ReisM. G.,
KoA. I.,
PicardeauM.2008; Targeted mutagenesis in pathogenic Leptospira species: disruption of the LigB gene does not affect virulence in animal models of leptospirosis. Infect Immun 76:5826–5833[CrossRef]
EllinghausenH. C.,
McCulloughW. G.1965; Nutrition of Leptospira pomona and growth of 13 other serotypes: fractionation of oleic albumin complex and a medium of bovine albumin and polysorbate 80. Am J Vet Res 26:45–51
FaisalS. M.,
YanW.,
ChenC. S.,
PalaniappanR. U.,
McDonoughS. P.,
ChangY. F.2008; Evaluation of protective immunity of Leptospira immunoglobulin like protein A (LigA) DNA vaccine against challenge in hamsters. Vaccine 26:277–287[CrossRef]
FaisalS. M.,
YanW.,
McDonoughS. P.,
ChangY. F.2009; Leptospira immunoglobulin-like protein A variable region (LigAvar) incorporated in liposomes and PLGA microspheres produces a robust immune response correlating to protective immunity. Vaccine 27:378–387[CrossRef]
GallowayR. L.,
LevettP. N.2008; Evaluation of a modified pulsed-field gel electrophoresis approach for the identification of Leptospira serovars. Am J Trop Med Hyg 78:628–632
HaakeD. A.,
SuchardM. A.,
KelleyM. M.,
DundooM.,
AltD. P.,
ZuernerR. L.2004; Molecular evolution and mosaicism of leptospiral outer membrane proteins involves horizontal DNA transfer. J Bacteriol 186:2818–2828[CrossRef]
HerrmannJ. L.,
BellengerE.,
PerolatP.,
BarantonG.,
GironsI. S.1992; Pulsed field gel electrophoresis of Not I digests of leptospiral DNA: a new rapid method of serovar identification. J Clin Microbiol 30:1696–1702
HookeyJ. V.,
BrydenJ.,
GatehouseL.1993; The use of 16S rDNA sequence analysis to investigate the phylogeny of Leptospiraceae and related spirochaetes. J Gen Microbiol 139:2585–2590[CrossRef]
JandaJ. M.,
AbbottS. L.2007; 16S rRNA gene sequencing for bacterial identification in the diagnostic laboratory: pluses, perils, and pitfalls. J Clin Microbiol 45:2761–2764[CrossRef]
La ScolaB.,
BuiL. T. M.,
BarantonG.,
KhamisA.,
RaoultD.2006; Partial rpoB gene sequencing for identification of Leptospira species. FEMS Microbiol Lett 263:142–147[CrossRef]
LevettP. N.2003; Usefulness of serologic analysis as a predictor of the infecting serovar in patients with severe leptospirosis. Clin Infect Dis 36:447–452[CrossRef]
LinY. P.,
ChangY. F.2007; A domain of the Leptospira LigB contributes to high affinity binding of fibronectin. Biochem Biophys Res Commun 362:443–448[CrossRef]
MajedZ.,
BellengerE.,
PosticD.,
PourcelC.,
BarantonG.,
PicardeauM.2005; Identification of variable-number tandem-repeat loci in Leptospira interrogans sensu stricto. J Clin Microbiol 43:539–545[CrossRef]
MatsunagaJ.,
BarocchiM. A.,
CrodaJ.,
YoungT. A.,
SanchezY.,
SiqueiraI.,
BolinC. A.,
ReisM. G.,
RileyL. W.other authors2003; Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily. Mol Microbiol 49:929–945[CrossRef]
MatsunagaJ.,
SanchezY.,
XuX.,
HaakeD. A.2005; Osmolarity, a key environmental signal controlling expression of leptospiral proteins LigA and LigB and the extracellular release of LigA. Infect Immun 73:70–78[CrossRef]
MatthiasM. A.,
RicaldiJ. N.,
CespedesM.,
DiazM. M.,
GallowayR. L.,
SaitoM.,
SteigerwaltA. G.,
PatraK. P.,
OreC. V.other authors2008; Human leptospirosis caused by a new, antigenically unique Leptospira associated with a Rattus species reservoir in the Peruvian Amazon. PLoS Negl Trop Dis 2:e213[CrossRef]
McBrideA. J. A.,
CerqueiraG. M.,
SuchardM. A.,
MoreiraA. N.,
ZuernerR. L.,
ReisM. G.,
HaakeD. A.,
KoA. I.,
DellagostinO. A.2009; Genetic diversity of the leptospiral immunoglobulin-like (Lig) genes in pathogenic Leptospira spp. Infect Genet Evol 9:196–205[CrossRef]
NascimentoA. L.,
KoA. I.,
MartinsE. A.,
Monteiro-VitorelloC. B.,
HoP. L.,
HaakeD. A.,
Verjovski-AlmeidaS.,
HartskeerlR. A.,
MarquesM. V.other authors2004; Comparative genomics of two Leptospira interrogans serovars reveals novel insights into physiology and pathogenesis. J Bacteriol 186:2164–2172[CrossRef]
PalaniappanR. U.,
ChangY. F.,
HassanF.,
McDonoughS. P.,
PoughM.,
BarrS. C.,
SimpsonK. W.,
MohammedH. O.,
ShinS.other authors2004; Expression of leptospiral immunoglobulin-like protein by Leptospira interrogans and evaluation of its diagnostic potential in a kinetic ELISA. J Med Microbiol 53:975–984[CrossRef]
PalaniappanR. U.,
ChangY. F.,
ChangC. F.,
PanM. J.,
YangC. W.,
HarpendingP.,
McDonoughS. P.,
DuboviE.,
DiversT.other authors2005; Evaluation of lig -based conventional and real time PCR for the detection of pathogenic leptospires. Mol Cell Probes 19:111–117
PerolatP.,
MerienF.,
EllisW. A.,
BarantonG.1994; Characterization of Leptospira isolates from serovars hardjo by ribotyping, arbitrarily primed PCR, and mapped restriction site polymorphisms. J Clin Microbiol 32:1949–1957
PosticD.,
Riquelme-SertourN.,
MerienF.,
PerolatP.,
BarantonG.2000; Interest of partial 16S rDNA gene sequences to resolve heterogeneities between Leptospira collections: application to L. meyeri
. Res Microbiol 151:333–341[CrossRef]
SalaünL.,
MérienF.,
GurianovaS.,
BarantonG.,
PicardeauM.2006; Application of multilocus variable-number tandem-repeat analysis for molecular typing of the agent of leptospirosis. J Clin Microbiol 44:3954–3962[CrossRef]
SilvaE. F.,
MedeirosM. A.,
McBrideA. J.,
MatsunagaJ.,
EstevesG. S.,
RamosJ. G.,
SantosC. S.,
CrodaJ.,
HommaA.other authors2007; The terminal portion of leptospiral immunoglobulin-like protein LigA confers protective immunity against lethal infection in the hamster model of leptospirosis. Vaccine 25:6277–6286[CrossRef]
SilvaE. F.,
SantosC. S.,
AthanazioD. A.,
SeyffertN.,
SeixasF. K.,
CerqueiraG. M.,
FagundesM. Q.,
BrodC. S.,
ReisM. G.other authors2008; Characterization of virulence of Leptospira isolates in a hamster model. Vaccine 26:3892–3896[CrossRef]
SlackA. T.,
DohntM. F.,
SymondsM. L.,
SmytheL. D.2005; Development of a multiple-locus variable number of tandem repeat analysis (MLVA) for Leptospira interrogans and its application to Leptospira interrogans serovar Australis isolates from Far North Queensland, Australia. Ann Clin Microbiol Antimicrob 4:10[CrossRef]
SlackA. T.,
SymondsM. L.,
DohntM. F.,
SmytheL. D.2006; Identification of pathogenic Leptospira species by conventional or real-time PCR and sequencing of the DNA gyrase subunit B encoding gene. BMC Microbiol 6:95[CrossRef]
SrimanoteP.,
WongdeethaiN.,
JieanampunkulP.,
SamonkiertS.,
LeepiyasakulchaiC.,
KalambahetiT.,
PrachayasittikulV.2008; Recombinant LigA for leptospirosis diagnosis and ligA among the Leptospira spp. clinical isolates. J Microbiol Methods 72:73–81[CrossRef]
VictoriaB.,
AhmedA.,
ZurnerR. L.,
AhmedN.,
BulachD. M.,
QuinteiroJ.,
HartskeerlR. A.2008; Conservation of the S10-spc - α locus within otherwise highly plastic genomes provides phylogenetic insight into the genus Leptospira
. PLoS One 3:e2752[CrossRef]
VijayachariP.,
AhmedN.,
SugunanA. P.,
GhousunnisaS.,
RaoK. R.,
HasnainS. E.,
SehgalS. C.2004; Use of fluorescent amplified fragment length polymorphism for molecular epidemiology of leptospirosis in India. J Clin Microbiol 42:3575–3580[CrossRef]
YanW.,
FaisalS. M.,
McDonoughS. P.,
DiversT. J.,
BarrS. C.,
ChangC. F.,
PanM. J.,
ChangY. F.2009; Immunogenicity and protective efficacy of recombinant Leptospira immunoglobulin-like protein B (rLigB) in a hamster challenge model. Microbes Infect 11:230–237[CrossRef]
YasudaP. H.,
SteigerwaltA. G.,
SulzerL. R.,
KaufmannA. F.,
RogersF.,
BrennerD. J.1987; Deoxyribonucleic acid relatedness between serogroups and serovars in the family Leptospiraceae with proposals for seven new Leptospira species. Int J Syst Bacteriol 37:407–415[CrossRef]
Distribution of the leptospiral immunoglobulin-like (lig) genes in pathogenic Leptospira species and application of ligB to typing leptospiral isolates