Chapter 55 : Serological and Molecular Diagnosis of Fungal Infections

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Analysis of the signs and symptoms of disease, in conjunction with an evaluation of available epidemiologic information and the results of modern imaging procedures, can often provide a presumptive clinical diagnosis of a fungal infection. However, the clinical presentation of many mycotic diseases is nonspecific, and a presumptive diagnosis must therefore be confirmed by appropriate laboratory tests. A definitive diagnosis of fungal disease is usually based upon the isolation of the etiologic agent in culture and/or microscopic demonstration of the organism in histopathologic or other clinical specimens. Unfortunately, these laboratory methods are insensitive and often unsuccessful, despite repeated sampling efforts. In the absence of positive microscopy or culture, immunologic and molecular tests offer alternative laboratory procedures to aid in the diagnosis of a mycotic disease. Advances in molecular diagnostic methods and the commercial introduction of new antigen and antibody detection tests hold promise for an earlier and more specific diagnosis than previously possible. Antibody detection tests are often helpful for the diagnosis of fungal infections. For example, when acute- and convalescent-phase serum specimens are tested, seroconversion from negative to positive or a 4-fold rise in antibody titer is considered to be diagnostic.

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55
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Image of FIGURE 1

Examples of ID results observed when a seven-well agar gel pattern is used. (A) Band of identity (patient 2); (B) band of partial identity (patient 1); (C) band of nonidentity (patient 3). Ag, reference antigen; Ab, reference antiserum; 1 to 4, patient test sera. A standard reference band is shown midway between the antiserum-containing wells and the central antigen-containing wells in all three panels.

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55
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Image of FIGURE 2

Diagram displaying organization of the microtiter plates and results for the CF assay. Solid circles, 100% lysis; shaded circles, 30 to 90% lysis; open circles, 0% lysis. SC, serum control wells receiving buffer in place of antigen; H, column receiving mycelial histoplasmin antigen; Y, column receiving yeast antigen; B, column receiving antigen; C, column receiving coccidioidin antigen; P, column receiving yeast antigen; CC, cell control; CH, amount of complement required for 50% lysis of SRBC. (A) Patient plate. Patient 1 demonstrates serum that is positive for antibodies with a 1:16 titer against mycelial histoplasmin antigen and a 1:64 titer against yeast antigen. Patient 2 displays serum that contains an AC titer of 1:16. However, because antibodies to coccidioidin are present at a titer that is >2 dilutions beyond the AC titer, the patient CF result is AC 1:16, 1:128. (B) Control plate. Columns 1 to 9 depict the reactivity of the control sera to specific fungal antigens. Columns 10 to 12, rows A to F, depict the results of the back titration of complement in CH units (5, 2.5, or 1.25) in the presence of fungal antigens or the VBD buffer control. Columns 11 and 12, row H, depict the cell control wells that receive SRBC only.

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55
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Generic image for table

Commercially available antibody detection tests for fungal diseases

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55
Generic image for table

Commercially available antigen detection tests for fungal diseases

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55
Generic image for table

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55
Generic image for table

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55
Generic image for table

Citation: Lindsley M. 2016. Serological and Molecular Diagnosis of Fungal Infections , p 503-534. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch55

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