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Chapter 114 : General Approaches for Direct Detection of Fungi

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Abstract:

This chapter on general approaches for direct detection of fungi reviews non-culture-based detection methods that can be performed directly with clinical materials in a clinical laboratory. These methods include direct microscopic examination, antigen detection, detection of fungus-specific metabolites, detection of cell wall components, detection of fungus-specific nucleic acids, and serologic diagnosis. Differentiating yeast from mold is important because yeast may represent normal colonization, while mold can indicate a deeper infectious process. Distinguishing colonization from disease complicates the diagnosis of candidiasis by antigen detection. Mannan, which is highly immunogenic, is one of the major cell wall components of species and is shed into the blood during infection. The glucuronoxylomannan component of the capsular polysaccharide is used for diagnosis of cryptococcosis. Strains of A, D, and AD serotypes are now classified as , while B and C serotype strains are classified as . Studies are in progress to determine the potential of direct identification of fungi in positive blood culture broths and patient samples. White et al. summarizes an update on the PCR standardization project. In this review it was found that the efficiency of the PCR was limited by the extraction procedures utilized and the compliance of some centers in the study.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114

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Fungal Infections
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Image of FIGURE 1
FIGURE 1

(row 1, left) Gram stain of BAL specimen showing gram-negative branching septate hyphae of . Magnification, approximately ×1,000.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 2
FIGURE 2

(row 1, right) Gram stain of skin biopsy with gram-variable branching hyphae of . Magnification, approximately ×1,000.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 3
FIGURE 3

(row 2, left) Gram stain of skin biopsy with gram-negative aseptate hyphae of the group. Magnification, approximately ×1,000.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 4
FIGURE 4

(row 2, right) Gram stain of sputum with showing gram-positive budding yeast cells with pseudohyphae.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 5
FIGURE 5

(row 3, left) Gram stain of spleen demonstrating ghost-like budding yeast-like cells with pseudohyphae. Magnification, approximately ×1,000.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 6
FIGURE 6

(row 3, right) Gram stain of vaginal secretions with gram-positive budding yeast cells of .

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 7
FIGURE 7

(row 4, left) Gram stain of skin abscess with showing gram-positive and gram-variable budding yeast cells of variable size surrounded by amorphous orange halos.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 8
FIGURE 8

(row 4, right) Calcofluor white stain of skin lesion showing typical yeast cells of spp. Magnification, approximately ×400.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 9
FIGURE 9

(row 1, left) Gomori methenamine silver (GMS) stain of BAL specimen showing dichotomously branching septate hyphae. Magnification, approximately ×400.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 10
FIGURE 10

(row 1, right) Calcofluor white stain of BAL specimen showing dichotomously branching septate hyphae. Magnification, approximately ×500.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 11
FIGURE 11

(row 2, left) GMS stain of skin biopsy with spp. showing ribbon-like hyphae. Magnification, approximately ×400.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 12
FIGURE 12

(row 2, right) Calcofluor white stain of spp. showing ribbon-like hyphae. Magnification, approximately ×500.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 13
FIGURE 13

(row 3, left) GMS stain of lung biopsy specimen showing with bulb formation at hyphal tip. Magnification, approximately ×1,000.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 14
FIGURE 14

(row 3, right) H&E stain of lung biopsy specimen showing dichotomously branching septate hyphae (shown at arrow). Magnification, approximately ×400.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 15
FIGURE 15

(row 1, left) Diff-Quik stain of CSF with . Magnification, approximately ×400.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 16
FIGURE 16

(row 1, right) India ink preparation of CSF showing encapsulated budding yeast cell of . Magnification, approximately ×400.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 17
FIGURE 17

(row 2, left) H&E stain of eumycotic mycetoma granule of . Magnification, approximately ×400.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 18
FIGURE 18

(row 2, right) Toluidine blue stain of BAL specimen showing cysts of .

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 19
FIGURE 19

(row 3, left) Calcofluor white stain of toenail specimen showing arthroconidia and hyphae of . Magnification, approximately ×200.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 20
FIGURE 20

(row 3, right) Sclerotic cells (copper penny) in tissue of chromoblastomycosis. Magnification, ×1,000.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 21
FIGURE 21

(row 1, left) GMS stain of lymph node showing characteristic cigar-shaped yeast cells of .

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 22
FIGURE 22

(row 1, right) Gram stain of abscess material showing large, broad-based, budding yeast cell with thick refractile wall characteristic of .

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 23
FIGURE 23

(row 2, left) GMS stain of lymph node showing blastoconidia of . Magnification, ×625.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 24
FIGURE 24

(row 2, right) H&E stain of . Large, round, thick-walled spherules (10 to 80 μm in diameter) filled with endospores (2 to 5 μm in diameter). Young spherules have a clear center with peripheral cytoplasm and a prominent thick wall.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 25
FIGURE 25

(row 3, left) Bright-field photomicrograph of showing multiple budding yeast cells resembling mariner's wheels. Magnification, ×1,590.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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Image of FIGURE 26
FIGURE 26

FIGURE 26 (row 3, right) Wright-Giemsa stain of BAL specimen showing characteristic fission yeast cells of .

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
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References

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Tables

Generic image for table
TABLE 1

Selection of clinical specimens for recovery of opportunistic fungal pathogens

Adapted from reference . The anatomic site of the fungal infection can be an indicator of what mycotic agent to suspect, but in an immunocompromised patient population, virtually any fungus can be an opportunistic pathogen.

Dermatophytes usually do not invade tissue but colonize only the outer layer of the skin.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
Generic image for table
TABLE 2

Characteristic fungal elements seen by direct examination of clinical specimens

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114
Generic image for table
TABLE 3

Methods and stains available for direct microscopic detection of fungi in clinical specimens

H&E, hematoxylin and eosin; PAS, periodic acid-Schiff.

Citation: Shea Y. 2011. General Approaches for Direct Detection of Fungi , p 1776-1792. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch114

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