Gradient diffusion susceptibility testing for Neisseria gonorrhoeae: an accurate alternative to agar dilution in high-MIC strains?

Abstract

Introduction: The correlation of antimicrobial susceptibility testing (AST) between agar dilution and gradient diffusion for Neisseria gonorrhoeae is not well established, especially in strains with high MICs.

Aim: The objective of this study was to evaluate the accuracy of gradient diffusion for N. gonorrhoeae .

Methods: Fifty strains of N. gonorrhoeae , all tested by the agar dilution method according to CLSI methods and confirmed to be genetically distinct using molecular typing (NG-MAST), were selected. Isolates with high MICs were targeted. Gradient diffusion was performed for ceftriaxone (CRO), cefixime (CFX), azithromycin (AZT), tetracycline (TET) and fosfomycin (FOS) using two different commercial antimicrobial strips on different culture media (a non-commercial GC agar base with 1 % defined growth supplement and two commercial media). The performance of agar gradient diffusion was assessed based on accuracy, using essential and category agreements (EA and CA).

Results: Essential and categorical agreement were over 90 % for CRO, CFX and AZT on the two commercial agar media tested. Category disagreements were seen for CFX and AZT, mostly just very major errors. For TET, EA ranged from 80 to 96 % and CA ranged from 38 to 76 %, most of the misclassifications being minor errors. Finally, EA for FOS ranged between 80 and 98 %.

Conclusion: Gradient diffusion is an accurate and acceptable alternative for CRO, CFX and AZT. Caution is advised when MICs are reported by gradient diffusion approach breakpoints because of the possibility of very major errors. The use of gradient diffusion is limited for TET because of the high rate of minor errors.

Keywords: Neisseria gonorrhoeae; antimicrobial susceptibility testing; gradient diffusion.