Table 1.
Primers for simple PCR/sequencing and nested PCR/sequencing for drug susceptibility testing in M. leprae used in this study.
Table 2.
Primers and probes of Custom TaqMan SNP genotyping assays for the folP1 loci for drug susceptibility testing in M. leprae.
Table 3.
Clinical characteristics of the leprosy patients enrolled in this study.
Fig 1.
DNA concentrations (ng/μl) resulting from M. leprae clinic specimens.
Table 4.
Specificity of nested PCR, and TaqMan SNP genotyping assay for drug resistance testing of Mycobacterium leprae.
Table 5.
Results of DNA sequencing and TaqMan SNP genotyping in the folP1, rpoB and gyrA genes of Mycobacterium leprae in DRDRs.
Table 6.
Comparison of the results of three molecular assays of drug resistance using clinical samples of leprosy patients.
Table 7.
folP, gyrA, and rpoB gene mutations related to drug resistance in isolates from leprosy patients in China.
Fig 2.
Sanger sequencing chromatograms indicated drug resistance loci in the folP1 Gene of M. leprae.
(A) 53ACC and 55CCC; (B) 53GCC; (C) 53ATC; (D) 55CTC; (E) 55CGC; (F) 53[A/G]CC and 55C[C/T]C; and [G] ACC53A[C/T][C/T], CGG54[C/G]GG and CCC55C[C/G]C in the folP1 gene. (A) Wildtype homozygous; (B-E) mutant homozygous; (F-G) mutant heterozygous.
Fig 3.
Multicomponent plot figures indicated drug resistance loci (CCC55CGC) in the folP1 Gene of M. leprae.
The DNA samples were genotyped using the Custom TaqMan SNP Genotyping Assay System. Major (also wildtype) alleles were detected by 2'-chloro-7'-phenyl-1,4-dichloro-6-carboxyfluorescein (VIC)-labeled probes (green) and minor alleles by 6-carboxyfluorescein (FAM)-labeled probes (blue). (A) Mutant homozygous G/G. (B) Mutant heterozygous C/G. (C) Wildtype homozygous C/C; (D) Allele discrimination plot showing alleles as wildtype homozygous C/C (lower right cluster), mutant heterozygous C/G (middle cluster), and mutant homozygous G/G (upper left) for the CCC55CGC loci of folP1 gene by Custom TaqMan SNP Genotyping Assay using 7500 Software v2.3, Applied Biosystem, USA. SNP indicates a single-nucleotide polymorphism.
Fig 4.
An allele discrimination plot indicated drug resistance loci in the folP1 Gene of M. leprae.
(A) ACC53GCC, (B) ACC53ATC, (C) CCC55CGC, and (D) CCC55CTC. Red, green, and blue dots represent homozygous genotypes, heterozygous genotypes and mutant homozygous genotypes, respectively, and the cross on the bottom left of the plot indicates the no-template control or failed PCR.
Table 8.
The mutation rates of new and relapsed leprosy patients.