Antifungal Susceptibility Testing, Therapy, and Prevention

Published on 08/02/2015 by admin

Filed under Basic Science

Last modified 22/04/2025

Print this page

rate 1 star rate 2 star rate 3 star rate 4 star rate 5 star
Your rating: none, Average: 5 (1 votes)

This article have been viewed 1949 times

Antifungal Susceptibility Testing, Therapy, and Prevention

Objectives

1. Name the documents available that contain the current guidelines for antifungal susceptibility testing.

2. Identify three circumstances in which antifungal susceptibility testing may be valuable.

3. List three areas of concern that complicate interpretive guidelines.

4. Explain how amphotericin B is produced, how it is administered, and its most significant adverse reaction.

5. Describe the mechanism of action of flucytosine and the drug’s therapeutic use.

6. Identify three echinocandins and describe their mechanism of action.

Antifungal Susceptibility Testing

Antifungal susceptibility tests are designed to provide information that helps the physician select the appropriate antifungal agent to treat a specific infection. Although antifungal susceptibility testing perhaps has not advanced as far as methods for determining the susceptibility of bacteria to antimicrobial agents, significant progress has been made. Substantial efforts have attempted to develop a standardized method that is reproducible among different laboratories. All of the technical variables in the testing process have been standardized, and efforts are underway to develop interpretative guidelines for different antifungal agents.

The Clinical Laboratory Standards Institute (CLSI) sets the standards for antifungal susceptibility testing. The current guidelines for these tests are provided in the following three documents, which are available on the CLSI website (www.clsi.org):

• Document M27-A3, Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, Approved Standard, 3rd edition. This document covers requirements for use of the broth microdilution method. The standards for susceptibility testing are very specific about the inoculum size, test medium, incubation time and temperature, and end point of yeasts that cause invasive fungal infections.

• Document M38-A2, Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi, Approved Standard, 2nd edition. This standard is a microdilution method for molds that cause invasive and cutaneous infections.

• Document M44-A2, Method for Antifungal Disk Diffusion Susceptibility Testing of Yeasts, 2nd edition, Approved Guideline. This standard provides methodology for disk diffusion testing for Candida spp., including quality control and interpretation guidelines. Also see the International Supplement M44S3.

It must be emphasized that the methodology and interpretation of antifungal susceptibility tests continue to evolve, and the laboratory should check for updated standards or a regular basis. Antifungal susceptibility tests are costly and time-consuming, but they may have value in the following circumstances:

The interpretative breakpoints for fluconazole, itraconazole, and flucytosine are based on experience in treating patients with mucosal infections, but they also appear to be consistent with information assembled for invasive infections. Problems that complicate the interpretative guidelines include:

Isolates of the same species may exhibit differences in minimum inhibitory concentrations (MICs) because of previous exposure to antifungal agents and/or acquisition of a genetic mechanism of resistance. For example, some isolates of Candida glabrata show susceptibility to fluconazole, but others do not. CLSI’s interpretative guidelines should be followed whenever possible, but anecdotal experience also is useful.

Despite the problems associated with antifungal susceptibility testing, many physicians believe that these tests are important for selecting an appropriate antifungal agent and as a method to detect the development of resistance of certain organisms during chemotherapy. A laboratory that is not equipped to perform CLSI methods, or validated equivalent methods, for susceptibility of clinically important fungal isolates should be prepared to send the isolate to a reference laboratory for testing. Amphotericin B, 5-fluorocytosine, ketoconazole, itraconazole, voriconazole, and fluconazole are common antifungal agents that traditionally have been tested; the newer antifungal agents may now be added to this list. For those interested in further information on this topic, the susceptibilities of many of the newer antifungal agents to some of the more challenging fungal pathogens have been published in a minireview by Lass-Florl et al.

Antifungal Therapy and Prevention

Numerous antifungal agents have been developed, and newer agents are on the horizon. The increasing number of immunosuppressed patients and the expansion of drug resistance of microorganisms make the development and appropriate use of antimicrobial agents two of the most important areas in microbiology and infectious diseases. This section is meant only to introduce the reader to the more commonly used antifungal agents; it is by no means comprehensive. Also, this section is not to be used as a guide for therapy. Therapeutic guidelines may be found in the texts listed in the bibliography.

Polyene Macrolide Antifungals

Polyene macrolide antifungal agents consist of a group of complex organic molecules, most of which contain multiple, conjugated, double-bond and one- to three-ring structures. This group includes many of the most commonly used antifungal agents, such as amphotericin B, the colloidal and liposomal preparations of amphotericin B, nystatin, and griseofulvin.

Amphotericin B

Amphotericin B is produced by the actinomycete Streptomyces nodosus. It is commonly infused intravenously to treat deep-seated fungal infections (e.g., invasive aspergillosis), and those caused by Candida spp., Cryptococcus spp., and members of the Mucorales. Amphotericin B binds the ergosterol component of the fungal cell membrane and alters the selective permeability of this membrane. However, other sterols, including those present in mammalian cell membranes, are also bound. The most significant adverse reaction associated with amphotericin B therapy is renal insufficiency. The liposomal amphotericin B compounds reportedly diminish this adverse reaction. Although amphotericin B is active against a wide variety of fungi, resistant organisms exist, which the laboratory must be able to identify. Fungi resistant to amphotericin B include Pseudallescheria boydii, Aspergillus terreus, Trichosporon spp., and in most cases Fusarium spp.

Azole Antifungal Drugs

The azole group of antifungal agents consists of the imidazoles and the triazoles. These compounds contain six carbon ring structures with conjugated double bonds, chloride residues, and five carbon ring structures that contain at least two nitrogen molecules. Traditionally used agents in this group include clotrimazole, miconazole, fluconazole, itraconazole, voriconazole and ketoconazole. The newer triazoles are voriconazole, posaconazole and, most recently, ravuconazole; of these only voriconazole is discussed here, because it was the first of the newer agents released and has been the most thoroughly reviewed. Azole antifungal agents disrupt the integrity of the fungal cell membrane by interfering with the synthesis of ergosterol.

Echinocandins

The echinocandins are glucan synthesis inhibitors. More specifically, they inhibit 1, 3 beta-glucan synthase, an enzyme important in fungal cell wall synthesis. These drugs lead to cellular osmotic instability. The three echinocandins are caspofungin, micafungin, and anidulafungin. Caspofungin was the first to be released and is the representative compound for this group.

Related posts:

Upper Respiratory Tract Infections and Other Infections of the Oral Cavity and Neck Specimen Management Erysipelothrix, Lactobacillus, and Similar Organisms Obligate Intracellular and Nonculturable Bacterial Agents Other Protozoa Haemophilus