Dermatophytosis laboratory findings: Difference between revisions
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=== Direct microscopic examination === | === Direct microscopic examination === | ||
* Treatment of skin specimen with 10–20 percent potassium hydroxide (KOH) is a convenient bedside tool to provide evidence of dermatophytic infection. | * Treatment of skin specimen with 10–20 percent potassium hydroxide (KOH) is a convenient bedside tool to provide evidence of dermatophytic infection.<ref name="pmid22474120">{{cite journal |vauthors=Kelly BP |title=Superficial fungal infections |journal=Pediatr Rev |volume=33 |issue=4 |pages=e22–37 |year=2012 |pmid=22474120 |doi=10.1542/pir.33-4-e22 |url=}}</ref> | ||
* Positive scrapings are characterized by: | * Positive scrapings are characterized by: | ||
** Refractile, long, smooth, undulating, branching, and septate hyphal filaments with or without arthroconidiospores. | ** Refractile, long, smooth, undulating, branching, and septate hyphal filaments with or without arthroconidiospores. | ||
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=== Culture and antifungal sensitivity === | === Culture and antifungal sensitivity === | ||
* Sabouraud dextrose agar (SDA, 4% peptone, 1% glucose, agar, water) is the most commonly used isolation media for dermatophytosis and serves as the medium for fungal growth. | * Sabouraud dextrose agar (SDA, 4% peptone, 1% glucose, agar, water) is the most commonly used isolation media for dermatophytosis and serves as the medium for fungal growth. | ||
* Development of colony takes 7–14 days. | * Development of colony takes 7–14 days.<ref name="pmid16479181">{{cite journal |vauthors=Ecemis T, Degerli K, Aktas E, Teker A, Ozbakkaloglu B |title=The necessity of culture for the diagnosis of tinea pedis |journal=Am. J. Med. Sci. |volume=331 |issue=2 |pages=88–90 |year=2006 |pmid=16479181 |doi= |url=}}</ref><ref name="pmid1379146">{{cite journal |vauthors=Rezabek GH, Friedman AD |title=Superficial fungal infections of the skin. Diagnosis and current treatment recommendations |journal=Drugs |volume=43 |issue=5 |pages=674–82 |year=1992 |pmid=1379146 |doi= |url=}}</ref><ref name="pmid22474120">{{cite journal |vauthors=Kelly BP |title=Superficial fungal infections |journal=Pediatr Rev |volume=33 |issue=4 |pages=e22–37 |year=2012 |pmid=22474120 |doi=10.1542/pir.33-4-e22 |url=}}</ref> | ||
* Modified | * Modified culturing techniques, with addition of gentamicin, chloramphenicol and cycloheximide is more selective for dermatophytes as chroramphenicol inhibits the growth of saprophytic fungus. | ||
* Dermatophyte test medium is an alternative to isolation media that contain pH indicator phenol red. It is incubated at room temperature for 5–14 days. | * Dermatophyte test medium is an alternative to isolation media that contain pH indicator phenol red. It is incubated at room temperature for 5–14 days. | ||
* Dermatophytes utilize the protein resulting in excess ammonium ion and alkaline environment which turn the medium from yellow to bright red. | * Dermatophytes utilize the protein resulting in excess ammonium ion and alkaline environment which turn the medium from yellow to bright red. | ||
=== Hematoxylin and eosin staining === | === Hematoxylin and eosin staining === | ||
* Histology may be used in diagnosis of Majocchi's granuloma in which KOH examination of scale may be false negative. | * Histology may be used in diagnosis of Majocchi's granuloma in which KOH examination of scale may be false negative.<ref name="pmid14744085">{{cite journal |vauthors=Al-Amiri A, Chatrath V, Bhawan J, Stefanato CM |title=The periodic acid-Schiff stain in diagnosing tinea: should it be used routinely in inflammatory skin diseases? |journal=J. Cutan. Pathol. |volume=30 |issue=10 |pages=611–5 |year=2003 |pmid=14744085 |doi= |url=}}</ref><ref name="pmid21987154">{{cite journal |vauthors=Bressan AL, Silva RS, Fonseca JC, Alves Mde F |title=Majocchi's granuloma |journal=An Bras Dermatol |volume=86 |issue=4 |pages=797–8 |year=2011 |pmid=21987154 |doi= |url=}}</ref> | ||
* Hyphae may be visualized in stratum corneum on hematoxylin and eosin staining. | * Hyphae may be visualized in stratum corneum on hematoxylin and eosin staining. | ||
* Special stains most commonly used are: | * Special stains most commonly used are: | ||
Line 29: | Line 29: | ||
=== Dermoscopy === | === Dermoscopy === | ||
* Tinea capitis may show:<ref name="pmid25471133">{{cite journal |vauthors=Lacarrubba F, Verzì AE, Micali G |title=Newly described features resulting from high-magnification dermoscopy of tinea capitis |journal=JAMA Dermatol |volume=151 |issue=3 |pages=308–10 |year=2015 |pmid=25471133 |doi=10.1001/jamadermatol.2014.3313 |url=}}</ref> | * Tinea capitis may show:<ref name="pmid25471133">{{cite journal |vauthors=Lacarrubba F, Verzì AE, Micali G |title=Newly described features resulting from high-magnification dermoscopy of tinea capitis |journal=JAMA Dermatol |volume=151 |issue=3 |pages=308–10 |year=2015 |pmid=25471133 |doi=10.1001/jamadermatol.2014.3313 |url=}}</ref><ref name="pmid25728878">{{cite journal |vauthors=Arrazola-Guerrero J, Isa-Isa R, Torres-Guerrero E, Arenas R |title=[Tinea capitis. Dermoscopic findings in 37 patients] |language=Spanish; Castilian |journal=Rev Iberoam Micol |volume=32 |issue=4 |pages=242–6 |year=2015 |pmid=25728878 |doi=10.1016/j.riam.2014.09.002 |url=}}</ref> | ||
** Comma hairs, which are slightly curved, fractured hair shafts | ** Comma hairs, which are slightly curved, fractured hair shafts | ||
** Corkscrew hair shave. | ** Corkscrew hair shave. | ||
Line 37: | Line 37: | ||
=== Polymerase chain reaction and nucleic acid sequence based amplification === | === Polymerase chain reaction and nucleic acid sequence based amplification === | ||
These tests not only help in the rapid and early diagnosis of infection but also help in determining drug resistance, and include: | These tests not only help in the rapid and early diagnosis of infection but also help in determining drug resistance, and include:<ref name="pmid23287391">{{cite journal |vauthors=Miyajima Y, Satoh K, Uchida T, Yamada T, Abe M, Watanabe S, Makimura M, Makimura K |title=Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes |journal=J. Dermatol. Sci. |volume=69 |issue=3 |pages=229–35 |year=2013 |pmid=23287391 |doi=10.1016/j.jdermsci.2012.11.589 |url=}}</ref><ref name="pmid27057486">{{cite journal |vauthors=Sahoo AK, Mahajan R |title=Management of tinea corporis, tinea cruris, and tinea pedis: A comprehensive review |journal=Indian Dermatol Online J |volume=7 |issue=2 |pages=77–86 |year=2016 |pmid=27057486 |pmc=4804599 |doi=10.4103/2229-5178.178099 |url=}}</ref><ref name="pmid25418736">{{cite journal |vauthors=Spiliopoulou A, Bartzavali C, Jelastopulu E, Anastassiou ED, Christofidou M |title=Evaluation of a commercial PCR test for the diagnosis of dermatophyte nail infections |journal=J. Med. Microbiol. |volume=64 |issue=Pt 1 |pages=25–31 |year=2015 |pmid=25418736 |doi=10.1099/jmm.0.079962-0 |url=}}</ref> | ||
* '''Uniplex PCR''' for direct dermatophyte detection in clinical samples: A PCR for the direct detection of dermatophytes in skin scales is available as in-house PCR-ELISA assay which separately identifies numerous dermatophyte species. | * '''Uniplex PCR''' for direct dermatophyte detection in clinical samples: A PCR for the direct detection of dermatophytes in skin scales is available as in-house PCR-ELISA assay which separately identifies numerous dermatophyte species. | ||
* '''Multiplex PCR''' for fungal detection in dermatophytes: Commercially available multiplex PCR tests enable simultaneous amplification of 21 dermatomycotic pathogens with subsequent DNA detection by means of agarose gel electrophoresis. | * '''Multiplex PCR''' for fungal detection in dermatophytes: Commercially available multiplex PCR tests enable simultaneous amplification of 21 dermatomycotic pathogens with subsequent DNA detection by means of agarose gel electrophoresis. |
Revision as of 22:10, 26 July 2017
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Laboratory Findings
Specimen collection
Scraping should be collected from active margin and transported in a presterilized black chart paper which keeps the specimen dry thus, preventing over growth of bacterial contaminants.
Direct microscopic examination
- Treatment of skin specimen with 10–20 percent potassium hydroxide (KOH) is a convenient bedside tool to provide evidence of dermatophytic infection.[1]
- Positive scrapings are characterized by:
- Refractile, long, smooth, undulating, branching, and septate hyphal filaments with or without arthroconidiospores.
- Fluorescent staining of the cell wall is the most sensitive method to microscopically detect fungi in skin scales as well as in specimens from nails and hair.
Culture and antifungal sensitivity
- Sabouraud dextrose agar (SDA, 4% peptone, 1% glucose, agar, water) is the most commonly used isolation media for dermatophytosis and serves as the medium for fungal growth.
- Development of colony takes 7–14 days.[2][3][1]
- Modified culturing techniques, with addition of gentamicin, chloramphenicol and cycloheximide is more selective for dermatophytes as chroramphenicol inhibits the growth of saprophytic fungus.
- Dermatophyte test medium is an alternative to isolation media that contain pH indicator phenol red. It is incubated at room temperature for 5–14 days.
- Dermatophytes utilize the protein resulting in excess ammonium ion and alkaline environment which turn the medium from yellow to bright red.
Hematoxylin and eosin staining
- Histology may be used in diagnosis of Majocchi's granuloma in which KOH examination of scale may be false negative.[4][5]
- Hyphae may be visualized in stratum corneum on hematoxylin and eosin staining.
- Special stains most commonly used are:
- Periodic acid-Schiff
- Gomori methanamine silver
Dermoscopy
- Tinea capitis may show:[6][7]
- Comma hairs, which are slightly curved, fractured hair shafts
- Corkscrew hair shave.
- Broken hair
- Tinea corporis may show:
- Involvement of vellus hair
Polymerase chain reaction and nucleic acid sequence based amplification
These tests not only help in the rapid and early diagnosis of infection but also help in determining drug resistance, and include:[8][9][10]
- Uniplex PCR for direct dermatophyte detection in clinical samples: A PCR for the direct detection of dermatophytes in skin scales is available as in-house PCR-ELISA assay which separately identifies numerous dermatophyte species.
- Multiplex PCR for fungal detection in dermatophytes: Commercially available multiplex PCR tests enable simultaneous amplification of 21 dermatomycotic pathogens with subsequent DNA detection by means of agarose gel electrophoresis.
References
- ↑ 1.0 1.1 Kelly BP (2012). "Superficial fungal infections". Pediatr Rev. 33 (4): e22–37. doi:10.1542/pir.33-4-e22. PMID 22474120.
- ↑ Ecemis T, Degerli K, Aktas E, Teker A, Ozbakkaloglu B (2006). "The necessity of culture for the diagnosis of tinea pedis". Am. J. Med. Sci. 331 (2): 88–90. PMID 16479181.
- ↑ Rezabek GH, Friedman AD (1992). "Superficial fungal infections of the skin. Diagnosis and current treatment recommendations". Drugs. 43 (5): 674–82. PMID 1379146.
- ↑ Al-Amiri A, Chatrath V, Bhawan J, Stefanato CM (2003). "The periodic acid-Schiff stain in diagnosing tinea: should it be used routinely in inflammatory skin diseases?". J. Cutan. Pathol. 30 (10): 611–5. PMID 14744085.
- ↑ Bressan AL, Silva RS, Fonseca JC, Alves Mde F (2011). "Majocchi's granuloma". An Bras Dermatol. 86 (4): 797–8. PMID 21987154.
- ↑ Lacarrubba F, Verzì AE, Micali G (2015). "Newly described features resulting from high-magnification dermoscopy of tinea capitis". JAMA Dermatol. 151 (3): 308–10. doi:10.1001/jamadermatol.2014.3313. PMID 25471133.
- ↑ Arrazola-Guerrero J, Isa-Isa R, Torres-Guerrero E, Arenas R (2015). "[Tinea capitis. Dermoscopic findings in 37 patients]". Rev Iberoam Micol (in Spanish; Castilian). 32 (4): 242–6. doi:10.1016/j.riam.2014.09.002. PMID 25728878.
- ↑ Miyajima Y, Satoh K, Uchida T, Yamada T, Abe M, Watanabe S, Makimura M, Makimura K (2013). "Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes". J. Dermatol. Sci. 69 (3): 229–35. doi:10.1016/j.jdermsci.2012.11.589. PMID 23287391.
- ↑ Sahoo AK, Mahajan R (2016). "Management of tinea corporis, tinea cruris, and tinea pedis: A comprehensive review". Indian Dermatol Online J. 7 (2): 77–86. doi:10.4103/2229-5178.178099. PMC 4804599. PMID 27057486.
- ↑ Spiliopoulou A, Bartzavali C, Jelastopulu E, Anastassiou ED, Christofidou M (2015). "Evaluation of a commercial PCR test for the diagnosis of dermatophyte nail infections". J. Med. Microbiol. 64 (Pt 1): 25–31. doi:10.1099/jmm.0.079962-0. PMID 25418736.