The NCCLS reference methodology for antifungal susceptibility testing is a new milestone of the evolution of medical mycology. The use of this methodology however, is not problem-free. At present, major limitations are a trailing phenomenon with azoles, unreliable detection of resistance to amphotericin B, poor growth of some organisms and unpractical procedures for the clinical laboratory. Herein a overview of NCCLS guidelines for yeasts and filamentous fungi is presented. Likewise, a review of studies conducted trying to overcome the limitations of reference procedures is also included. Several alternative approaches are reviewed as alternative media, inoculum size and incubation time. Modifications of reading procedure and endpoint determination are also evaluated. Agar diffusion methods and other methods for susceptibility testing are cited. Finally, we discuss the data on correlation of the in vitro results with the in vivo activity.