TABLE 2.

Spoligotyping and rifoligotyping patterns and sequence analysis for DNA obtained from rifampin-resistant M. tuberculosis isolates and the corresponding ZN-stained slidesa

Sample no.CountrySpoligotyping patternRifoligotyping patternrpoB mutationScore for ZN-stained smear on Bronk- horst scale
StrainZNStrainZNStrainZN
R1Moldova▪▪▪▪▪▪▪□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□▪□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□▪□□□□▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+2
R2Moldova▪▪▪▪▪▪▪□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□▪□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□▪□□□□▪▪▪▪▪▪▪▪□▪▪▪ □□□□▪□▪▪▪ □□□□516-GTC516-GTC+2
R3Moldova▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪□ □▪□□▪▪▪▪□ □▪□□531-TGG531-TGG+5
R4Moldova□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+5
R5Moldova□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪□▪▪▪ □□□□▪▪▪▪□ ▪□□□516-TTC531-TTG+5
R6Moldova□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪□▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+5
R7Moldova▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪□ □▪□□▪▪▪▪□ □▪□□531-TGG531-TGG+2
R8Moldova□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□Neg531-TTGNegNeg
R9Moldova▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+2
R10The Netherlands□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪Neg▪□▪▪▪ □□□□Neg516-GTCNegNeg
R11The Netherlands▪▪▪▪▪▪▪□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□▪□□□□▪▪▪▪▪▪▪Neg▪▪▪▪□ □▪□□Neg531-TGGNegNeg
R12The Netherlands▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪▪□□□□□□□□□□▪▪▪▪▪▪▪▪□□□□▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+2
R13The NetherlandsNo strain available□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪ND▪▪▪□▪ □□□▪ND526-TAC+1
R14Kazakhstan□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪▪ □□□□▪▪▪▪▪ □□□□516-GTC516-GTC+2
R15Kazakhstan□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+3
R16Kazakhstan□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+2
R17Kazakhstan□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+2
R18Kazakhstan□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪▪▪▪▪□ ▪□□□▪▪▪▪□ ▪□□□531-TTG531-TTG+2
R19KazakhstanNo strain available□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪ND▪▪▪▪□ ▪□□□ND531-TTG+1
R20KazakhstanNo strain available□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪ND▪▪▪▪□ ▪□□□ND531-TTG+1
R21KazakhstanNo strain available□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□▪▪▪▪▪▪▪▪▪NDNegNDNeg+1
  • a Characteristics of the rifampin-resistant strains that were isolated (strain) and extracts from ZN-stained slides (ZN) for different patients from Moldova, The Netherlands, and Kazakhstan. Spoligotyping patterns are read from left to right and indicate spacers 1 to 43 from the direct repeat region, as described by Kamerbeek et al. (15). A fragment of the rpoB gene was amplified from bacterial cultures and ZN-stained slides and tested for mutations by hybridization to membrane-bound oligonucleotides giving rifoligotyping patterns corresponding to alleles for sensitivity (squares 1 to 5; probes W1 to W5, respectively) and resistance alleles (squares 6 to 9; probes M1 to M4, respectively). ▪, hybridization; □, no hybridization; Neg, negative; ND, not determined.