Fungal Testing Overview

Fungal antigens, fungal DNA, and anti-fungal antibodies can be used as non-invasive biomarkers for invasive fungal infections (IFIs). MiraVista Diagnostics (MVD) specializes in developing diagnostic tests to detect these biomarkers. These tests provide a rapid, cost-efficient alternative to invasive testing, often required otherwise to diagnose IFIs. Over the past 20 years MVD tests have revolutionized diagnosis and treatment monitoring of IFIs in animals. Use has been reported in many peer-reviewed publications. The reported diagnostic performances are found below.

Diagnostic performance of MVD fungal tests. Bolded tests are the recommended first non-invasive diagnostic test(s) to be performed.

Pathogen Test Species Sample Sensitivity Specificity# Min. Vol. (ml)^ Test Code Ref.
Blastomyces* Antigen EIA K9 Urine 93 98 0.5 316 1Motschenbacher LO, Furrow E, Rendahl AK, et al. Retrospective analysis of the effects of Blastomyces antigen concentration in urine and radiographic findings on survival in dogs with blastomycosis. J Vet Intern Med 2021.,2Mourning AC, Patterson EE, Kirsch EJ, et al. Evaluation of an enzyme immunoassay for antibodies to a recombinant Blastomyces adhesin-1 repeat antigen as an aid in the diagnosis of blastomycosis in dogs. J Am Vet Med Assoc 2015;247:1133-1138.,3Spector D, Legendre AM, Wheat J, et al. Antigen and antibody testing for the diagnosis of blastomycosis in dogs. J Vet Intern Med 2008;22:839-843.,4Foy DS, Trepanier LA, Kirsch EJ, et al. Serum and urine Blastomyces antigen concentrations as markers of clinical remission in dogs treated for systemic blastomycosis. J Vet Intern Med 2014;28:305-310.
K9 Serum 91 100 0.8 316 2Mourning AC, Patterson EE, Kirsch EJ, et al. Evaluation of an enzyme immunoassay for antibodies to a recombinant Blastomyces adhesin-1 repeat antigen as an aid in the diagnosis of blastomycosis in dogs. J Am Vet Med Assoc 2015;247:1133-1138.,3Spector D, Legendre AM, Wheat J, et al. Antigen and antibody testing for the diagnosis of blastomycosis in dogs. J Vet Intern Med 2008;22:839-843.,5Arambulo PV, 3rd, Topacio TM, Jr., Famatiga EG, et al. Leptospirosis among abattoir employees, dog pound workers, and fish inspectors in the city of Manila. The Southeast Asian journal of tropical medicine and public health 1972;3:212-220.
Antibody EIA K9 Serum 82 97 0.25 330 2Mourning AC, Patterson EE, Kirsch EJ, et al. Evaluation of an enzyme immunoassay for antibodies to a recombinant Blastomyces adhesin-1 repeat antigen as an aid in the diagnosis of blastomycosis in dogs. J Am Vet Med Assoc 2015;247:1133-1138.,3Spector D, Legendre AM, Wheat J, et al. Antigen and antibody testing for the diagnosis of blastomycosis in dogs. J Vet Intern Med 2008;22:839-843.
Antibody ID K9 Serum 54 100 0.25 322 2Mourning AC, Patterson EE, Kirsch EJ, et al. Evaluation of an enzyme immunoassay for antibodies to a recombinant Blastomyces adhesin-1 repeat antigen as an aid in the diagnosis of blastomycosis in dogs. J Am Vet Med Assoc 2015;247:1133-1138.,3Spector D, Legendre AM, Wheat J, et al. Antigen and antibody testing for the diagnosis of blastomycosis in dogs. J Vet Intern Med 2008;22:839-843.,6Arceneaux KA, Taboada J, Hosgood G. Blastomycosis in dogs: 115 cases (1980-1995). J Am Vet Med Assoc 1998;213:658-664.,7Crews LJ, Feeney DA, Jessen CR, et al. Utility of diagnostic tests for and medical treatment of pulmonary blastomycosis in dogs: 125 cases (1989- 2006). J Am Vet Med Assoc 2008;232:222-227.
Histoplasma Antigen EIA K9 Urine 92 99 0.5 310 8Cunningham L, Cook A, Hanzlicek A, et al. Sensitivity and Specificity of Histoplasma Antigen Detection by Enzyme Immunoassay. J Am Anim Hosp Assoc 2015;51:306-310.,9Clark K, Hanzlicek AS. Evaluation of a novel monoclonal antibody-based enzyme immunoassay for detection of Histoplasma antigen in urine of dogs. J Vet Intern Med 2020.
Feline Urine 94 98 0.5 310 10Cook AK, Cunningham LY, Cowell AK, et al. Clinical evaluation of urine Histoplasma capsulatum antigen measurement in cats with suspected disseminated histoplasmosis. J Feline Med Surg 2012;14:512-515.,11Rothenburg L, Hanzlicek AS, Payton ME. A monoclonal antibody-based urine Histoplasma antigen enzyme immunoassay (IMMY(R)) for the diagnosis of histoplasmosis in cats. J Vet Intern Med 2019;33:603-610.
Feline Serum 73 NA 0.8 310 12Hanzlicek AS, Meinkoth JH, Renschler JS, et al. Antigen Concentrations as an Indicator of Clinical Remission and Disease Relapse in Cats with Histoplasmosis. J Vet Intern Med 2016;30:1065-1073.
Antibody EIA K9 Serum 76 93 0.25 327 MVD
Feline Serum 83 95 0.25 328 MVD
Antibody ID K9 Serum 28 100 0.25 321 MVD
Feline Serum 5 100 0.25 321 MVD
Coccidioides (valley fever) Antigen EIA Feline Urine 8 100 0.5 315 13Holbrook ED, Greene RT, Rubin SI, et al. Novel canine anti-Coccidioides immunoglobulin G enzyme immunoassay aids in diagnosis of coccidioidomycosis in dogs. Medical mycology 2019;57:800-806.,14Kirsch EJ, Greene RT, Prahl A, et al. Evaluation of Coccidioides antigen detection in dogs with coccidioidomycosis. Clin Vaccine Immunol 2012;19:343-345.
Feline Serum 27 97 0.8 315 13Holbrook ED, Greene RT, Rubin SI, et al. Novel canine anti-Coccidioides immunoglobulin G enzyme immunoassay aids in diagnosis of coccidioidomycosis in dogs. Medical mycology 2019;57:800-806.,14Kirsch EJ, Greene RT, Prahl A, et al. Evaluation of Coccidioides antigen detection in dogs with coccidioidomycosis. Clin Vaccine Immunol 2012;19:343-345.
Antibody EIA K9 Serum 89 94 0.25 329 13Holbrook ED, Greene RT, Rubin SI, et al. Novel canine anti-Coccidioides immunoglobulin G enzyme immunoassay aids in diagnosis of coccidioidomycosis in dogs. Medical mycology 2019;57:800-806.
Antibody ID K9 Serum 87 92 0.25 320 13Holbrook ED, Greene RT, Rubin SI, et al. Novel canine anti-Coccidioides immunoglobulin G enzyme immunoassay aids in diagnosis of coccidioidomycosis in dogs. Medical mycology 2019;57:800-806.,15Johnson LR, Herrgesell EJ, Davidson AP, et al. Clinical, clinicopathologic, and radiographic findings in dogs with coccidioidomycosis: 24 cases (1995-2000). J Am Vet Med Assoc 2003;222:461-466.,16Gunstra A, Steurer JA, Seibert RL, et al. Sensitivity of Serologic Testing for Dogs Diagnosed with Coccidioidomycosis on Histology: 52 Cases (2012- 2013). J Am Anim Hosp Assoc 2019;55:238-242.
Feline Serum 99 NA 0.25 320 17Arbona N, Butkiewicz CD, Keyes M, et al. Clinical features of cats diagnosed with coccidioidomycosis in Arizona, 2004-2018. J Feline Med Surg 2020;22:129-137,18Greene RT, Troy GC. Coccidioidomycosis in 48 cats: a retrospective study (1984-1993). J Vet Intern Med 1995;9:86-91.
Antibody ID+EIA K9 Serum 99 95 0.5 329
320
13Holbrook ED, Greene RT, Rubin SI, et al. Novel canine anti-Coccidioides immunoglobulin G enzyme immunoassay aids in diagnosis of coccidioidomycosis in dogs. Medical mycology 2019;57:800-806.
Cryptococcus Antigen LA K9 Serum 91 100 0.5 319 19Malik R, McPetrie R, Wigney DI, et al. A latex cryptococcal antigen agglutination test for diagnosis and monitoring of therapy for cryptococcosis. Aust Vet J 1996;74:358-364.,20Johnston L, Mackay B, King T, et al. Abdominal cryptococcosis in dogs and cats: 38 cases (2000-2018). J Small Anim Pract 2021;62:19-27.,21Trivedi SR, Sykes JE, Cannon MS, et al. Clinical features and epidemiology of cryptococcosis in cats and dogs in California: 93 cases (1988-2010). J Am Vet Med Assoc 2011;239:357-369.
Feline Serum 98 NA 0.25 319 19Malik R, McPetrie R, Wigney DI, et al. A latex cryptococcal antigen agglutination test for diagnosis and monitoring of therapy for cryptococcosis. Aust Vet J 1996;74:358-364.,21Trivedi SR, Sykes JE, Cannon MS, et al. Clinical features and epidemiology of cryptococcosis in cats and dogs in California: 93 cases (1988-2010). J Am Vet Med Assoc 2011;239:357-369.,22Medleau L, Marks MA, Brown J, et al. Clinical evaluation of a cryptococcal antigen latex agglutination test for diagnosis of cryptococcosis in cats. J Am Vet Med Assoc 1990;196:1470-1473.
Aspergillus (systemic) Antigen EIA K9 Urine 88 92 0.8 309 23Garcia RS, Wheat LJ, Cook AK, et al. Sensitivity and specificity of a blood and urine galactomannan antigen assay for diagnosis of systemic aspergillosis in dogs. J Vet Intern Med 2012;26:911-919.
K9 Serum 92 84 0.8 309 23Garcia RS, Wheat LJ, Cook AK, et al. Sensitivity and specificity of a blood and urine galactomannan antigen assay for diagnosis of systemic aspergillosis in dogs. J Vet Intern Med 2012;26:911-919.
K9 Ur+Ser 100 84-92 0.8 309 23Garcia RS, Wheat LJ, Cook AK, et al. Sensitivity and specificity of a blood and urine galactomannan antigen assay for diagnosis of systemic aspergillosis in dogs. J Vet Intern Med 2012;26:911-919.
Aspergillus (sinonasal) Antigen EIA K9 Serum 24 82 0.8 309 24Billen F, Peeters D, Peters IR, et al. Comparison of the value of measurement of serum galactomannan and Aspergillus-specific antibodies in the diagnosis of canine sino-nasal aspergillosis. Vet Microbiol 2009;133:358-365.
Feline Serum 23 78 0.8 309 25Whitney J, Beatty JA, Martin P, et al. Evaluation of serum galactomannan detection for diagnosis of feline upper respiratory tract aspergillosis. Vet Microbiol 2013;162:180-185.
Antibody EIA K9 Serum 76 100 0.25 324 24Billen F, Peeters D, Peters IR, et al. Comparison of the value of measurement of serum galactomannan and Aspergillus-specific antibodies in the diagnosis of canine sino-nasal aspergillosis. Vet Microbiol 2009;133:358-365.
Feline Serum 43 100 0.25 324 26Barrs VR, Ujvari B, Dhand NK, et al. Detection of Aspergillus-specific antibodies by agar gel double immunodiffusion and IgG ELISA in feline upper respiratory tract aspergillosis. Vet J 2015;203:285-289
For tests with multiple references the combined sensitivity and specificity was provided.
*Blastomyces antigen test on urine is also the first-choice test for cats.
^Minimum sample volume required for testing.
#Specificity was often provided for non-fungal disease.
EIA, enzyme immunoassay; ID, immunodiffusion; LA, latex agglutination

Geographic location of enzootic invasive fungal infections (IFIs) is an important consideration. Cryptococcosis and aspergillosis, along with other invasive molds, occur anywhere in the U.S. Although infections can occur sporadically anywhere, IFIs due to dimorphic fungi (Blastomyces, Histoplasma, and Coccidioides) are found more commonly in predictable locations. The map below provides a rough estimate of enzootic regions.

A Fungi Map from CDC Website

Invasive fungal infections (IFIs) commonly include non-specific findings such as lethargy, anorexia, weight loss, and fever unresponsive to antibiotics. Other more specific findings are also often present and can help guide diagnostic testing. Any IFI can affect any single organ or be multi-systemic, but patterns exist.

Common anatomic locations and clinical signs of invasive fungal infections in dogs and cats.

PATHOGEN SPECIES AFFECTED COMMON ORGANS INVOLVED COMMON CLINICAL SIGNS
Blastomyces Dog>>>Cat Lung
Lymph nodes
Skin
Eyes/periocular tissues
Bones/joints
Testicles/prostate (dog)
Anorexia
Weight loss
Fever
Tachypnea
Cough
Lameness
Vision disturbance
Cutaneous Lesions
Histoplasma Cat>Dog Lung
Lymph node
Spleen & Liver
Eyes/periocular tissues
Bone marrow
Bone/joints (cat)
GI tract (dog)
Anorexia
Weight loss
Fever
Tachypnea
Lameness
Vision disturbance
Cutaneous lesions
Diarrhea
Cryptococcus Cat>>>Dog Nasal cavity & sinuses
Eyes & periocular tissues
Lymph nodes
Skin
CNS
Abdominal viscera (dog)
Nasal discharge
Facial deformity
Cutaneous lesions
Blindness
CNS signs
Vomiting, diarrhea (dog)
Coccidioides
(valley fever)
Dog>>Cat Lung
Lymph nodes
Skin
Eyes
CNS
Bones/joints
Anorexia
Weight loss
Tachypnea
Cough
Lameness
Cutaneous lesions
CNS signs
Aspergillus
(systemic)
Dog>>>Cat Bone/joints
Vertebral end plates
CNS
Kidney
Lymph nodes
Skin
Eyes
Abdominal viscera
Anorexia
Weight loss
Fever
Lameness
Vision disturbance
CNS signs
Cutaneous lesions
PU/PD
Aspergillus
(sinonasal)
Dog>>Cat Nasal cavity
Sinuses
Orbit
CNS
Nasal discharge
Nasal depigmentation
Facial deformity
CNS signs

With many available tests, choosing the best test can be challenging. In some cases, a combination of tests for a specific pathogen is preferred. For others, sequential testing might be needed. Considering the overlap of clinical findings, and enzootic areas, testing for multiple fungal pathogens is appropriate in many cases. To help simplify test selection, and treatment considerations, algorithms are provided below.

Diagnostic and Treatment Algorithm for Blastomycosis in Dogs or Cats

Diagnostic and Treatment Algorithm for Histoplasmosis in Dogs or Cats

Diagnostic and Treatment Algorithm for Coccidioidomycosis (Valley Fever) in Dogs or Cats

Diagnostic and Treatment Algorithm for Cryptococcosis in Dogs or Cats

Diagnostic and Treatment Algorithm for Systemic Aspergillosis in Dogs or Cats

Diagnostic and Treatment Algorithm for Sinonasal Aspergillosis in Dogs or Cats

REFERENCES:

  1. Motschenbacher LO, Furrow E, Rendahl AK, et al. Retrospective analysis of the effects of Blastomyces antigen concentration in urine and radiographic findings on survival in dogs with blastomycosis. J Vet Intern Med 2021.
  2. Mourning AC, Patterson EE, Kirsch EJ, et al. Evaluation of an enzyme immunoassay for antibodies to a recombinant Blastomyces adhesin-1 repeat antigen as an aid in the diagnosis of blastomycosis in dogs. J Am Vet Med Assoc 2015;247:1133-1138.
  3. Spector D, Legendre AM, Wheat J, et al. Antigen and antibody testing for the diagnosis of blastomycosis in dogs. J Vet Intern Med 2008;22:839-843.
  4. Foy DS, Trepanier LA, Kirsch EJ, et al. Serum and urine blastomyces antigen concentrations as markers of clinical remission in dogs treated for systemic blastomycosis. J Vet Intern Med 2014;28:305-310.
  5. Arambulo PV, 3rd, Topacio TM, Jr., Famatiga EG, et al. Leptospirosis among abattoir employees, dog pound workers, and fish inspectors in the city of Manila. The Southeast Asian journal of tropical medicine and public health 1972;3:212-220.
  6. Arceneaux KA, Taboada J, Hosgood G. Blastomycosis in dogs: 115 cases (1980-1995).. J Am Vet Med Assoc 1998;213:658-664.
  7. Crews LJ, Feeney DA, Jessen CR, et al. Utility of diagnostic tests for and medical treatment of pulmonary blastomycosis in dogs: 125 cases (1989- 2006). J Am Vet Med Assoc 2008;232:222-227.
  8. Cunningham L, Cook A, Hanzlicek A, et al. Sensitivity and Specificity of Histoplasma Antigen Detection by Enzyme Immunoassay. J Am Anim Hosp Assoc 2015;51:306-310.
  9. Clark K, Hanzlicek AS. Evaluation of a novel monoclonal antibody-based enzyme immunoassay for detection of Histoplasma antigen in urine of dogs. J Vet Intern Med 2020.
  10. Cook AK, Cunningham LY, Cowell AK, et al. Clinical evaluation of urine Histoplasma capsulatum antigen measurement in cats with suspected disseminated histoplasmosis. J Feline Med Surg 2012;14:512-515.
  11. Rothenburg L, Hanzlicek AS, Payton ME. A monoclonal antibody-based urine Histoplasma antigen enzyme immunoassay (IMMY(R)) for the diagnosis of histoplasmosis in cats. J Vet Intern Med 2019;33:603-610.
  12. Hanzlicek AS, Meinkoth JH, Renschler JS, et al. Antigen Concentrations as an Indicator of Clinical Remission and Disease Relapse in Cats with Histoplasmosis. J Vet Intern Med 2016;30:1065-1073.
  13. Holbrook ED, Greene RT, Rubin SI, et al. Novel canine anti-Coccidioides immunoglobulin G enzyme immunoassay aids in diagnosis of coccidioidomycosis in dogs Medical mycology 2019;57:800-806.
  14. Kirsch EJ, Greene RT, Prahl A, et al. Evaluation of Coccidioides antigen detection in dogs with coccidioidomycosis. Clin Vaccine Immunol 2012;19:343-345.
  15. Johnson LR, Herrgesell EJ, Davidson AP, et al. Clinical, clinicopathologic, and radiographic findings in dogs with Coccidioidomycosis: 24 cases (1995-2000). J Am Vet Med Assoc 2003;222:461-466.
  16. Gunstra A, Steurer JA, Seibert RL, et al. Sensitivity of Serologic Testing for Dogs Diagnosed with Coccidioidomycosis on Histology: 52 Cases (2012- 2013). J Am Anim Hosp Assoc 2019;55:238-242.
  17. Arbona N, Butkiewicz CD, Keyes M, et al. Clinical features of cats diagnosed with coccidioidomycosis in Arizona, 2004-2018 J Feline Med Surg 2020;22:129-137.
  18. Greene RT, Troy GC. Coccidioidomycosis in 48 cats: a retrospective study (1984-1993) J Vet Intern Med 1995;9:86-91.
  19. Malik R, McPetrie R, Wigney DI, et al. A latex cryptococcal antigen agglutination test for diagnosis and monitoring of therapy for cryptococcosis. Aust Vet J 1996;74:358-364.
  20. Johnston L, Mackay B, King T, et al. Abdominal cryptococcosis in dogs and cats: 38 cases (2000-2018). J Small Anim Pract 2021;62:19-27.
  21. Trivedi SR, Sykes JE, Cannon MS, et al. Clinical features and epidemiology of cryptococcosis in cats and dogs in California: 93 cases (1988-2010). J Am Vet Med Assoc 2011;239:357-369.
  22. Medleau L, Marks MA, Brown J, et al. Clinical evaluation of a cryptococcal antigen latex agglutination test for diagnosis of cryptococcosis in cats. J Am Vet Med Assoc 1990;196:1470-1473.
  23. Garcia RS, Wheat LJ, Cook AK, et al. Sensitivity and specificity of a blood and urine galactomannan antigen assay for diagnosis of systemic aspergillosis in dogs. J Vet Intern Med 2012;26:911-919.
  24. Billen F, Peeters D, Peters IR, et al. Comparison of the value of measurement of serum galactomannan and Aspergillus-specific antibodies in the diagnosis of canine sino-nasal aspergillosis. Vet Microbiol 2009;133:358-365.
  25. Whitney J, Beatty JA, Martin P, et al. Evaluation of serum galactomannan detection for diagnosis of feline upper respiratory tract aspergillosis. Vet Microbiol 2013;162:180-185.
  26. Barrs VR, Ujvari B, Dhand NK, et al. Detection of Aspergillus-specific antibodies by agar gel double immunodiffusion and IgG ELISA in feline upper respiratory tract aspergillosis. Vet J 2015;203:285-289