Molecular typing of Leptospira spp. based on putative O-antigen polymerase gene (wzy), the benefit over 16S rRNA gene sequence.
Molecular typing of leptospiral strains based on variation within putative O-antigen polymerase gene (wzy) was determined among reference strains and those isolated from patients. Using the PCR primers designed from the flanking gene of wzy derived from Leptospira interrogans serovar Copenhageni, all L. interrogans serovars as well as human and rodent leptospiral isolates from Thailand could be amplified. The size of PCR product ranged from 1 to 1.5 kb. The limitation of these primer pairs was the inability to amplify those strains whose sequences differ in the region of the primers, these included Leptospira biflexa (serovar Patoc), Leptospira borgpetersenii (serovar Tarassovi) and Leptospira kirschneri (serovar Bim, Bulgarica, Butembo). Notably, amplification was not limited to L. interrogans as demonstrated by the amplification of some strains from L. kirschneri, Leptospira meyeri, Leptospira noguchii, Leptospira santarosai, L. borgpetersenii and Leptospira weilii. The phylogenetic tree of wzy sequence, inferred by posterior probability of the Bayesian, enabled the categorization of leptospiral serovars into seven genetically related group, of which its differentiation power was better than that of the more highly conserved 16S rRNA gene, which is used extensively for genotyping.
Wangroongsarb
Piyada
P
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Chanket
Teerarut
T
Gunlabun
Kata
K
Long
Do Hoang
DH
Satheanmethakul
Pedcharat
P
Jetanadee
Siriporn
S
Thaipadungpanit
Janjira
J
Wuthiekanun
Vannaporn
V
Peacock
Sharon J
SJ
Blacksell
Stuart D
SD
Smythe
Lee D
LD
Bulach
Dieter M
DM
Kalambaheti
Thareerat
T
D000818:Animals; D005838:Genotype; D006602:Hexosyltransferases / Q000235:genetics*; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D007922:Leptospirosis / Q000382:microbiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013785:Thailand
D012336:RNA, Ribosomal, 16S; D006602:Hexosyltransferases; C085652:O-antigen polymerase