Chapter 41 : Diagnostic Approach Based on Capsular Antigen, Capsule Detection, β-Glucan, and DNA Analysis

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The polysaccharide capsule is a unique feature of that distinguishes the organism from other medically important yeasts. Various histochemical stains react with cryptococci, including mucin stains such as mucicarmine, alcian blue, and periodic acid-Schiff that stain the capsule and Fontana-Masson stain, which interacts with the cell wall. The sensitivity of cerebrospinal fluid (CSF) antigen detection is highest for diagnosing cryptococcal meningitis in HIV-infected persons and varies based upon factors such as the fungal burden in the CSF and the presence of well-encapsulated organisms. Since the serum cryptococcal antigen is positive in almost all AIDS patients with cryptococcal meningitis, there has been interest in using the test to screen high-risk HIV-infected persons, such as those with fevers, neurological symptoms, and/or low CD4 counts. At the current time, there are four commercially available assays that detect β-Glucan (BG) in serum. Over the past two decades, a myriad of molecular techniques have been explored for the detection of pathogenic fungi. Molecular fingerprinting techniques, e.g., randomly amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), pulsed-field gel electrophoresis, amplified fragment length polymorphism (AFLP), PCR fingerprinting with minisatellite (M13) or microsatellite primers [e.g., (GACA) or (CTG)], karyotypes, sequencing, multilocus microsatellite typing, mating-type locus, and multilocus sequence typing (MLST), are among the most popular typing techniques that have been applied to discern and characterize the genetic heterogeneity within the species of the species complex.

Citation: Diaz M, Nguyen M. 2011. Diagnostic Approach Based on Capsular Antigen, Capsule Detection, β-Glucan, and DNA Analysis, p 547-564. In Heitman J, Kozel T, Kwon-Chung K, Perfect J, Casadevall A (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555816858.ch41
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Image of FIGURE 1

A schematic representation of the rRNA gene family organization. Different sets of primers can be used to amplify target regions such as the DI/D2 domain and the ITS and IGS regions. Arrows indicate the direction in which a given primer amplifies.

Citation: Diaz M, Nguyen M. 2011. Diagnostic Approach Based on Capsular Antigen, Capsule Detection, β-Glucan, and DNA Analysis, p 547-564. In Heitman J, Kozel T, Kwon-Chung K, Perfect J, Casadevall A (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555816858.ch41
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Generic image for table

Comparison of selected commercially available LA and EIA assays for detection of cryptococcal antigens

Citation: Diaz M, Nguyen M. 2011. Diagnostic Approach Based on Capsular Antigen, Capsule Detection, β-Glucan, and DNA Analysis, p 547-564. In Heitman J, Kozel T, Kwon-Chung K, Perfect J, Casadevall A (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555816858.ch41
Generic image for table

Molecular type concordance of C. and C.

Citation: Diaz M, Nguyen M. 2011. Diagnostic Approach Based on Capsular Antigen, Capsule Detection, β-Glucan, and DNA Analysis, p 547-564. In Heitman J, Kozel T, Kwon-Chung K, Perfect J, Casadevall A (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555816858.ch41

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