Article Figures & Data
Figures
- Fig. 1.
Detection of gyrA mutations by PCR-RFLP analysis (a) and PCR-SSCP analysis of SacII site-positive (b) and -negative (c) DNA. Lanes M, molecular size standard (HaeIII-digested φX174); lane +, SacII site-positive DNA fragment; lane −, SacII site-negative DNA fragment; lanes 1, wild type; lane 2, mutant containing alteration of GCC→GCT at codon 67 (silent); lane 3, mutant containing alteration of CGT→CGA at codon 68 (silent); lane 4, mutant containing alteration of CCG→CCA at codon 79 (silent); lane 5, mutant containing alteration of CGC→CGT at codon 91 (silent); lane 6, mutant containing alteration of Asp-87→Asn; lane 7, mutant containing alterations of CCG→CCA at codon 79 (silent) plus Asp-87→Gly; lane 8, mutant containing alterations of Ala-67→Ser plus Asp-87→Gly; lane 9, mutant containing alteration of Gln-106→Leu; lane 10, mutant containing alteration of Thr-83→Ile; lane 11, mutant containing alterations of CGT→CGA at codon 68 (silent) plus Thr-83→Ile; lane 12, mutant containing alterations of Thr-83→Ile plus CGC→CGT at codon 91 (silent); lane 13, mutant containing alterations of Thr-83→Ile plus Asp-87→Gly; lane 14, mutant containing alterations of Thr-83→Ile plus Asp-87→Asn; lane 15, mutant containing alterations of Thr-83→Ile plus Asp-87→His; lane 16, mutant containing alteration of GCG→GCA at codon 84 (silent); lane 17, mutant containing alteration of Ala-84→Pro; lane 18, mutant containing alterations of GCG→GCA at codon 84 (silent) plus Asp-87→Tyr. Lanes 6 through 13, 17, and 18, quinolone-resistant clinical strains; lanes 14 and 15, mutated plasmids derived from pGL2B5; lanes 16 and 18, generous gifts from T. Schollaardt, namely, P4394 and Y4492, respectively.
Tables
- Table 1.
Types of point mutation(s) and respective incidence among the 335 clinical isolates
SacII site Type of mutation Amino acid change Nucleotide change Incidence (no. [%] of strains) Positive Wild type None None 243 (72.5) Silent mutations Ala-67→Ala (silent) GCC→GCT 2 (0.6) Arg-68→Arg (silent) CGT→CGA 9 (2.7) Pro-79→Pro (silent) CCG→CCA 6 (1.8) Arg-91→Arg (silent) CGC→CGT 5 (1.5) Single missense mutations with or without silent mutation Asp-87→Asn GAC→AAC 2 (0.6) Pro-79, Asp-87→Pro (silent), Gly CCG, GAC→CCA, GGC 1 (0.3) Gln-106→Leu CAG→CTG 1 (0.3) Double missense mutations Ala-67, Asp-87→Ser, Gly GCC, GAC→TCC, GGC 1 (0.3) Negative Silent mutation Ala-84→Ala (silent) GCG→GCA 0a Single missense mutations with or without silent mutation Thr-83→Ile ACC→ATC 60 (17.9) Arg-68, Thr-83→Arg (silent), Ile CGT, ACC→CGA, ATC 2 (0.6) Thr-83, Arg-91→Ile, Arg (silent) ACC, CGC→ATC, CGT 1 (0.3) Ala-84→Pro GCG→CCG 1 (0.3) Ala-84, Asp-87→Ala (silent), Tyr GCG, GAC→GCA, TAC 0b Double missense mutations Thr-83, Asp-87→Ile, Gly ACC, GAC→ATC, GGC 1 (0.3) Thr-83, Asp-87→Ile, Asn ACC, GAC→ATC, AAC 0c Thr-83, Asp-87→Ile, His ACC, GAC→ATC, CAC 0c Total 335 (100) - Table 2.
Susceptibilities of 336 clinical isolates of P. aeruginosa to 10 fluoroquinolones
Drug MIC (μg/ml) for strains with the following GyrA alterationsa (type of change): PAO1 Wild type (n = 265)b Thr-83→Ile (PU→NH) (n = 63) Ala-84→Pro (NH→NH) (n = 1) Asp-87→Gly (−1c→PU) (n = 1) Asp-87→Asn (−1→PU) (n = 2) Asp-87→Tyrd (−1→PU) (n = 1) Gln-106→Leu (PU→NH) (n = 1) Ala-67, Asp-87→Ser, Gly (NH, −1→PU, PU) (n = 1) Thr-83, Asp-87→Ile, Gly (PU, −1→NH, PU) (n = 1) Range 50% 90% Range 50% 90% Norfloxacin 0.39 0.20–100 0.78 3.13 1.56–400 100 200 25 12.5 1.56, 12.5 12.5 25 12.5 200 Ofloxacin 0.78 0.39–100 1.56 6.25 3.13–800 50 200 25 12.5 3.13, 25 12.5 25 12.5 200 Enoxacin 0.78 0.20–200 1.56 6.25 3.13–>400 100 >400 25 25 3.13, 25 50 25 25 200 Ciprofloxacin 0.10 0.025–50 0.20 0.78 0.39–400 25 50 6.25 3.13 0.78, 3.13 6.25 3.13 3.13 50 Tosufloxacin 0.20 0.05–>25 0.39 1.56 0.10–>25 >25 >25 6.25 6.25 0.78, 1.56 3.13 1.56 6.25 >25 Lomefloxacin 0.78 0.39–200 1.56 6.25 0.78–800 200 400 50 25 6.25, 50 25 50 25 400 Fleroxacin 0.78 0.20–200 1.56 6.25 3.13–>1,600 100 400 25 12.5 3.13, 25 25 25 12.5 400 Sparfloxacin 0.78 0.20–200 1.56 3.13 1.56–>1,600 100 200 12.5 12.5 1.56, 12.5 12.5 25 6.25 200 Gatifloxacin 0.78 0.20–50 1.56 3.13 1.56–200 50 100 12.5 6.25 1.56, 6.25 6.25 12.5 6.25 50 CS-940 0.39 0.10–50 0.78 3.13 1.56–800 50 100 6.25 3.13 0.78, 12.5 3.13 12.5 6.25 100
