A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans

JoAnna Rudasill; Sheila Criswell

doi:doi:10.11648/j.ijidt.20230801.12

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A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans

JoAnna Rudasill, Sheila Criswell

Published in International Journal of Infectious Diseases and Therapy (Volume 8, Issue 1)

Received: 30 December 2022 Accepted: 1 February 2023 Published: 9 February 2023

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Abstract

Background: Using special stains to detect fungus is an essential diagnostic tool in histology. Because microbiology cultures can take over a month for a definitive diagnosis, rapid detection and treatment of fungal infections are necessary for patient care, particularly in immunocompromised patients when infections may lead to life-threatening complications. Objective: A comparison of various classic special stain methods along with more recent modifications on those staining methods was performed to evaluate the effectiveness of fungal detection while attempting to ameliorate the dangers associated with certain chemicals used in histochemical procedures. Methods: Twenty-five formalin-fixed paraffin-embedded archived tissue blocks containing the genera Aspergillus, Blastomyces, Candida, Cryptococcus, Histoplasma, Mucor, and Pneumocystis were stained with periodic acid Schiff light green (PAS-LG) and Grocott methenamine silver (GMS) using standard methods. Modifications of these and other protocols found in the literature to include altering the oxidizing reagent, time in oxidizer, temperature of oxidizer, pretreatment, and counterstain applied were made and the results were compared. Results: Cryptococcus and Candida were the most readily labeled fungal genera with detection rates of 92% and 85%, respectively across all staining protocols assayed. Histoplasma, however proved to be the most difficult genus to successfully label with only a 21% sensitivity across all staining protocols. Conclusion: While several procedures which included altering oxidizers, incubation periods, and temperatures were variably successful, the original Grocott method with chromic acid as the oxidizer remained the most effective protocol in the detection of fungal elements with 96% sensitivity.

Published in	International Journal of Infectious Diseases and Therapy (Volume 8, Issue 1)
DOI	10.11648/j.ijidt.20230801.12
Page(s)	10-22
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2023. Published by Science Publishing Group

Keywords

Fungus, GMS, Silver Stain, Periodic Acid Schiff

References

[1]	Youngberg G, Wallen E, Giorgadze T. Narrow-spectrum histochemical staining of fungi. Arch. Pathol. Lab. Med. 2003; 127: 1529–1530.
[2]	Shimono L & Hartman B. A simple and reliable rapid methenamine silver stain for Pneumocystis carinii and fungi. Archives of Pathology and Laboratory Medicine. 1986; 110 (9): 855–856.
[3]	Broadwater D, Messersmith L, Bishop B, Tomkovich A, Salinas J, Lynch C. Development and Validation of Ultra-Rapid Periodic Acid–Schiff Stain for Use in Identifying Fungus on Frozen Section. Arch Pathol Lab Med 2022; doi: https://doi.org/10.5858/arpa.2021-0273-OA
[4]	Kligman, A. M., Meson, H. and DeLamater, E. D. The Hotchkiss-McManus stain for the histopathologic diagnosis of fungus diseases. Am. J. Clin. Pathol. 1951; 21: 86–91.
[5]	Grocott R. A stain for fungi in tissue sections and smears using Gomori’s methenamine-silver nitrate technic. American Journal of Clinical Pathology. 1955; 25: 975-979.
[6]	Gridley M. A stain for fungi in tissue sections. Am. J. Pathol. 1953; 23: 303–307.
[7]	Gomori, G. A new histochemical test for glycogen and mucin. Am. J. Pathol. 1946: 10: 177–179.
[8]	Carson F, Fredenburgh J, Maxwell J. Inconsistent detection of Histoplasma capsulatum with periodic acid oxidation in the Grocott methenamine-silver nitrate (GMS) fungus stain. Journal of Histotechnology, 1999; 22: 2, 119-122.
[9]	PubChem, 2022 https://pubchem.ncbi.nlm.nih.gov/compound/Chromic-acid#section=Toxicity
[10]	Shiogama K, Kitazawa K, Mizutani Y, Onouchi T, Inada K, Tsutsumi Y. New Grocott stain without using chromic acid. Acta Histochem. Cytochem. 2015; 48 (1): 9-14.
[11]	Ku N, Pullarkat S, Kim Y, Cheng L, O’Malley D. Use of Cd42b immunohistochemical stain for the detection of Histoplasma. Annals of Diagnostic Pathology. 2018; 32: 47-50.
[12]	Stanley M, Henry M, Iber C. Foamy alveolar casts: diagnostic specificity for Pneumocystis carinii pneumonia in bronchoalveolar lavage fluid cytology. Diagn Cytopathol. 1988; 4 (2): 113-5.
[13]	Almeida M, Almeida-Silva F, Guimaraes A, Almeida-Paes R, Zancope-Oliviera R. The occurrence of histoplasmosis in Brazil: A systematic review. International Journal of Infectious Diseases. 2019; 86: 147-156.
[14]	Lockhart S and Guarner J. Emerging and reemerging fungal infections. Seminars in Diagnostic Pathology. 2019; 36 (3): 177-181. https://doi.org/10.1053/j.semdp.2019.04.010.
[15]	Pincelli T, Enzler M, Davis M, Tande A, Comfere N, Bruce A. Oropharyngeal histoplasmosis: a report of 10 cases. Clinical and Experimental Dermatology, 2019. doi: 10.1111/ced.13927.

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APA Style

JoAnna Rudasill, Sheila Criswell. (2023). A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans. International Journal of Infectious Diseases and Therapy, 8(1), 10-22. https://doi.org/10.11648/j.ijidt.20230801.12

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ACS Style

JoAnna Rudasill; Sheila Criswell. A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans. Int. J. Infect. Dis. Ther. 2023, 8(1), 10-22. doi: 10.11648/j.ijidt.20230801.12

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AMA Style

JoAnna Rudasill, Sheila Criswell. A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans. Int J Infect Dis Ther. 2023;8(1):10-22. doi: 10.11648/j.ijidt.20230801.12

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@article{10.11648/j.ijidt.20230801.12,
  author = {JoAnna Rudasill and Sheila Criswell},
  title = {A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans},
  journal = {International Journal of Infectious Diseases and Therapy},
  volume = {8},
  number = {1},
  pages = {10-22},
  doi = {10.11648/j.ijidt.20230801.12},
  url = {https://doi.org/10.11648/j.ijidt.20230801.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijidt.20230801.12},
  abstract = {Background: Using special stains to detect fungus is an essential diagnostic tool in histology. Because microbiology cultures can take over a month for a definitive diagnosis, rapid detection and treatment of fungal infections are necessary for patient care, particularly in immunocompromised patients when infections may lead to life-threatening complications. Objective: A comparison of various classic special stain methods along with more recent modifications on those staining methods was performed to evaluate the effectiveness of fungal detection while attempting to ameliorate the dangers associated with certain chemicals used in histochemical procedures. Methods: Twenty-five formalin-fixed paraffin-embedded archived tissue blocks containing the genera Aspergillus, Blastomyces, Candida, Cryptococcus, Histoplasma, Mucor, and Pneumocystis were stained with periodic acid Schiff light green (PAS-LG) and Grocott methenamine silver (GMS) using standard methods. Modifications of these and other protocols found in the literature to include altering the oxidizing reagent, time in oxidizer, temperature of oxidizer, pretreatment, and counterstain applied were made and the results were compared. Results: Cryptococcus and Candida were the most readily labeled fungal genera with detection rates of 92% and 85%, respectively across all staining protocols assayed. Histoplasma, however proved to be the most difficult genus to successfully label with only a 21% sensitivity across all staining protocols. Conclusion: While several procedures which included altering oxidizers, incubation periods, and temperatures were variably successful, the original Grocott method with chromic acid as the oxidizer remained the most effective protocol in the detection of fungal elements with 96% sensitivity.},
 year = {2023}
}

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TY  - JOUR
T1  - A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans
AU  - JoAnna Rudasill
AU  - Sheila Criswell
Y1  - 2023/02/09
PY  - 2023
N1  - https://doi.org/10.11648/j.ijidt.20230801.12
DO  - 10.11648/j.ijidt.20230801.12
T2  - International Journal of Infectious Diseases and Therapy
JF  - International Journal of Infectious Diseases and Therapy
JO  - International Journal of Infectious Diseases and Therapy
SP  - 10
EP  - 22
PB  - Science Publishing Group
SN  - 2578-966X
UR  - https://doi.org/10.11648/j.ijidt.20230801.12
AB  - Background: Using special stains to detect fungus is an essential diagnostic tool in histology. Because microbiology cultures can take over a month for a definitive diagnosis, rapid detection and treatment of fungal infections are necessary for patient care, particularly in immunocompromised patients when infections may lead to life-threatening complications. Objective: A comparison of various classic special stain methods along with more recent modifications on those staining methods was performed to evaluate the effectiveness of fungal detection while attempting to ameliorate the dangers associated with certain chemicals used in histochemical procedures. Methods: Twenty-five formalin-fixed paraffin-embedded archived tissue blocks containing the genera Aspergillus, Blastomyces, Candida, Cryptococcus, Histoplasma, Mucor, and Pneumocystis were stained with periodic acid Schiff light green (PAS-LG) and Grocott methenamine silver (GMS) using standard methods. Modifications of these and other protocols found in the literature to include altering the oxidizing reagent, time in oxidizer, temperature of oxidizer, pretreatment, and counterstain applied were made and the results were compared. Results: Cryptococcus and Candida were the most readily labeled fungal genera with detection rates of 92% and 85%, respectively across all staining protocols assayed. Histoplasma, however proved to be the most difficult genus to successfully label with only a 21% sensitivity across all staining protocols. Conclusion: While several procedures which included altering oxidizers, incubation periods, and temperatures were variably successful, the original Grocott method with chromic acid as the oxidizer remained the most effective protocol in the detection of fungal elements with 96% sensitivity.
VL  - 8
IS  - 1
ER  -

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Author Information

JoAnna Rudasill

Department of Diagnostic and Health Sciences, University of Tennessee Health Science Center, Memphis, USA
Sheila Criswell

Department of Diagnostic and Health Sciences, University of Tennessee Health Science Center, Memphis, USA

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APA Style

JoAnna Rudasill, Sheila Criswell. (2023). A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans. International Journal of Infectious Diseases and Therapy, 8(1), 10-22. https://doi.org/10.11648/j.ijidt.20230801.12

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ACS Style

JoAnna Rudasill; Sheila Criswell. A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans. Int. J. Infect. Dis. Ther. 2023, 8(1), 10-22. doi: 10.11648/j.ijidt.20230801.12

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AMA Style

JoAnna Rudasill, Sheila Criswell. A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans. Int J Infect Dis Ther. 2023;8(1):10-22. doi: 10.11648/j.ijidt.20230801.12

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@article{10.11648/j.ijidt.20230801.12,
  author = {JoAnna Rudasill and Sheila Criswell},
  title = {A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans},
  journal = {International Journal of Infectious Diseases and Therapy},
  volume = {8},
  number = {1},
  pages = {10-22},
  doi = {10.11648/j.ijidt.20230801.12},
  url = {https://doi.org/10.11648/j.ijidt.20230801.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijidt.20230801.12},
  abstract = {Background: Using special stains to detect fungus is an essential diagnostic tool in histology. Because microbiology cultures can take over a month for a definitive diagnosis, rapid detection and treatment of fungal infections are necessary for patient care, particularly in immunocompromised patients when infections may lead to life-threatening complications. Objective: A comparison of various classic special stain methods along with more recent modifications on those staining methods was performed to evaluate the effectiveness of fungal detection while attempting to ameliorate the dangers associated with certain chemicals used in histochemical procedures. Methods: Twenty-five formalin-fixed paraffin-embedded archived tissue blocks containing the genera Aspergillus, Blastomyces, Candida, Cryptococcus, Histoplasma, Mucor, and Pneumocystis were stained with periodic acid Schiff light green (PAS-LG) and Grocott methenamine silver (GMS) using standard methods. Modifications of these and other protocols found in the literature to include altering the oxidizing reagent, time in oxidizer, temperature of oxidizer, pretreatment, and counterstain applied were made and the results were compared. Results: Cryptococcus and Candida were the most readily labeled fungal genera with detection rates of 92% and 85%, respectively across all staining protocols assayed. Histoplasma, however proved to be the most difficult genus to successfully label with only a 21% sensitivity across all staining protocols. Conclusion: While several procedures which included altering oxidizers, incubation periods, and temperatures were variably successful, the original Grocott method with chromic acid as the oxidizer remained the most effective protocol in the detection of fungal elements with 96% sensitivity.},
 year = {2023}
}

Copy | Download

TY  - JOUR
T1  - A Comparison of Histological Staining Methods for Pathogenic Fungi in Humans
AU  - JoAnna Rudasill
AU  - Sheila Criswell
Y1  - 2023/02/09
PY  - 2023
N1  - https://doi.org/10.11648/j.ijidt.20230801.12
DO  - 10.11648/j.ijidt.20230801.12
T2  - International Journal of Infectious Diseases and Therapy
JF  - International Journal of Infectious Diseases and Therapy
JO  - International Journal of Infectious Diseases and Therapy
SP  - 10
EP  - 22
PB  - Science Publishing Group
SN  - 2578-966X
UR  - https://doi.org/10.11648/j.ijidt.20230801.12
AB  - Background: Using special stains to detect fungus is an essential diagnostic tool in histology. Because microbiology cultures can take over a month for a definitive diagnosis, rapid detection and treatment of fungal infections are necessary for patient care, particularly in immunocompromised patients when infections may lead to life-threatening complications. Objective: A comparison of various classic special stain methods along with more recent modifications on those staining methods was performed to evaluate the effectiveness of fungal detection while attempting to ameliorate the dangers associated with certain chemicals used in histochemical procedures. Methods: Twenty-five formalin-fixed paraffin-embedded archived tissue blocks containing the genera Aspergillus, Blastomyces, Candida, Cryptococcus, Histoplasma, Mucor, and Pneumocystis were stained with periodic acid Schiff light green (PAS-LG) and Grocott methenamine silver (GMS) using standard methods. Modifications of these and other protocols found in the literature to include altering the oxidizing reagent, time in oxidizer, temperature of oxidizer, pretreatment, and counterstain applied were made and the results were compared. Results: Cryptococcus and Candida were the most readily labeled fungal genera with detection rates of 92% and 85%, respectively across all staining protocols assayed. Histoplasma, however proved to be the most difficult genus to successfully label with only a 21% sensitivity across all staining protocols. Conclusion: While several procedures which included altering oxidizers, incubation periods, and temperatures were variably successful, the original Grocott method with chromic acid as the oxidizer remained the most effective protocol in the detection of fungal elements with 96% sensitivity.
VL  - 8
IS  - 1
ER  -

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