pseudomallei MSHR840, 7 – B thailandensis 82172, 8 – B thailand

pseudomallei MSHR840, 7 – B. thailandensis 82172, 8 – B. thailandensis-like MSMB122, 9 – B. ubonensis MSMB108, 10 – Burkholderia sp. MSMB175, 11 – B. thailandensis-like MSMB43. Lanes 1–3 are representative of type A strains, Lanes 4–5 are representative of type

B strains, Lanes 6–10 are representative of type B2 strains, and Lane 11 contains an unknown serotype B O-antigen Twenty-one DNA Damage inhibitor strains of B. mallei expressed type A O-antigen while the remaining two strains (ATCC10399 and NCTC120) expressed rough type. ATCC10399 was previously described as having an intact ladder [13, 20], but the whole genome sequence (WGS) available in GenBank shows an IS407A insertion in wbiG (NZ_CH899681), which would predict a rough type. IS407A is known as one of the most common insertion sequence (IS) elements in B. pseudomallei and B. mallei[21]. NCTC120’s rough type phenotype is consistent with prior works [13, 20]. Further immunoblotting with the B. mallei LPS-specific mAb 3D11

showed all 21 B. mallei strains with intact ladder profiles bound this antibody while the two rough type strains did not. B. pseudomallei K96243 and B. oklahomensis E0147 bound mAb 3D11, as previously described [11]. Similarly, eight of the B. thailandensis strains bound mAb 3D11 while E264, MSMB59, MSMB60, and 82172 did not (Additional file 1: Table S1). Similarly, testing the strains containing type A with the IgM mAb Pp-PS-W, the B. pseudomallei LPS-specific mAb [13], showed that B. mallei ATCC23344 and B. oklahomensis Selleck Staurosporine E0147 were not seroreactive while B. pseudomallei K96243 was seroreactive. Notably, nine B. thailandensis strains were seroreactive to this mAb, while MSMB59 and MSMB60 were not. This suggested the existence of seroreactivity Adenosine triphosphate diversity within B. thailandensis. PCR suggested that 11 strains of B. ubonensis would be positive for type B O-antigen. Immunoblotting confirmed the expression of type B in all of these, one of which, MSMB57, was selected for genomic analysis. Another strain, B. ubonensis MSMB108, was negative for all genotypes by PCR but displays

a ladder pattern identical to the type B2 B. thailandensis-like MSMB122 (Figure 1). We also noted that other tested B. ubonensis strains produced distinct LPS ladder patterns to those of B. pseudomallei, which were not seroreactive (data not shown). Along with B. thailandensis, B. ubonensis was the only species that expressed more than one type of B. pseudomallei O-antigen. B. thailandensis-like strains expressed two different O-antigen ladder patterns, both of which were B serotypes. Strains MSMB121, 122, 712, and 714 expressed ladder type B2 (Additional file 1: Table S1), whereas strain MSMB43 expressed a novel serologically related O-antigen not found in B. pseudomallei. This O-antigen, like type B2, bound the type B patient’s serum but exhibited a banding pattern unlike either type B or B2 (Figure 1).

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