Tariq Butt, Muhammad Yasin Khan, Rifat Nadeem Ahmad, Muhammad Salman, Raja Kamran Afzal.
Validity of Nalidixic acid screening in Fluoroquinolone-resistant Typhoid Salmonellae.
J Coll Physicians Surg Pak Jan ;16(1):31-4.

Objective: To validate the screening of low-level fluoroquinolone resistance in typhoid salmonellae by using nalidixic acid (30mg) disk providing an acceptable zone of inhibition. Design: Quasi-experimental study. Place and Duration of Study: The Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, Pakistan from July 2002 to June 2003. Materials and Methods: Antimicrobial susceptibility of 225 clinical isolates of S. typhi (n=126) and S. paratyphi A (n=99) against nalidixic acid and ciprofloxacin was determined by the modified Kirby-Bauer disk diffusion and agar dilution techniques of NCCLS. The relationship between the zone sizes and the MICs of the two quinolones was plotted in the form of scattergrams and nalidixic acid MICs and zone of inhibition sizes were correlated with those of ciprofloxacin by regression analysis. Results: One hundred and ninety-five isolates were nalidixic acid-susceptible (MIC <16µg/mL) and ~30 were nalidixic acid-resistant (MIC >32 µg/mL). All the nalidixic acid-susceptible isolates had ciprofloxacin MIC of <0.064µg/mL. Among the nalidixic acid-resistant isolates ~20 had ciprofloxacin MIC >0.125µg/mL and ~10 had ciprofloxacin MIC <0.03- 0.064µg/mL. The diameter of inhibition zone around a 30mg nalidixic acid disk of nalidixic acid-resistant isolates was <13 mm (range 6-16 mm, mean 10.3 mm + SD 3.5 mm), while among nalidixic acid-susceptible isolates it ranged from 14 to 30 mm (mean 23.8 mm + SD 2.2 mm). The diameter of inhibition zone around a 5mg ciprofloxacin disk of nalidixic acidresistant isolates ranged from 26 to 35 mm (mean 29.8 mm + SD 3.1 mm), while in nalidixic acid-susceptible isolates it ranged from 32 to 42 mm (mean 36.6 mm + SD 1.9 mm). With ciprofloxacin MIC Ž0.125 mg/mL taken as a breakpoint, a zone of <33mm around a 5mg ciprofloxacin disk to detect low susceptibility strains had a sensitivity of 100% and a specificity of 82%. Screening for nalidixic acid resistance (inhibition zone diameter of <13 mm) in isolates with ciprofloxacin MIC Ž 0.125mg/mL using a 30mg nalidixic acid disk yielded a sensitivity of 100% and a specificity of 95%. Conclusion: Screening for nalidixic acid resistance with a 30mg nalidixic acid disk is a reliable and cost-effective method for detection of low-level fluoroquinolone resistance in typhoid salmonellae.

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