Molecular Diagnostics of Histoplasmosis

Histoplasmosis is a systemic fungal disease that occurs worldwide. The highest incidence of the disease is reported on the American continent. It also occurs in China, India, South-Eastern Asia, Africa, Australia and Europe. Clinical syndromes of histoplasmosis are not specific and in most cases immunocompetent individuals are asymptomatic or present mild influenza-like disease. Immunocompromised patients especially individuals with AIDS, can develop a severe and fatal disease due to fungal dissemination to many organs. Etiological agent of histoplasmosis is the dimorphic fungus Histoplasma capsulatum, which inhabits the soils contaminated with bird or bat droppings. Three biological varieties are considered for this fungus: H. capsulatum var. capsulatum, H. capsulatum var. duboissii and H. capsulatum var. farciminosum. Genetic differences are observed among H. capsulatum strains from diverse regions of the world. The main molecular methodologies for genetic typing of fungi are based on DNA fingerprinting. They have been an important instrument to identify possible sources of infection in outbreaks of histoplasmosis. Genetic profiles of H. capsulatum, isolated from bats and humans, helped to understand the distribution of the disease in certain endemic regions. The con-ventional diagnosis of histoplasmosis is performed by means of cultural and microscopic examination of samples from the respiratory tract and biologic fluids. However, these techniques yield positive results in only 50 % of cases. In the last two decades, approaches for the detecting of H. capsulatum in clinical samples, using different molecular targets, based on PCR assay have been developed. Their use can shorten the time span of analysis for diagnosis confirmation. Molecular methods have high specificity and sensitivity and reduce the risk of infection for the laboratory personnel. In this study we reviewed the recently published data on the use of main molecular methods for diagnosis of histoplasmosis.

1. Wheat L.J., Azar M.M., Bahr N.C., Spec A., Relich R.F., Hage C. Histoplasmosis. Infect. Dis. Clin. N. Am. 2016; 30(1):207— 27. DOI: 10.1016/i.idc.2015.10.009.

2. Negroni R. Histoplasmosis en America Latina. Biomedica: revista del Instituto National de Salud. 2011; 31(3):301—4. DOI: 10.7705/biomedica.v31i3.597.

3. Colombo A.L., Tobon A., Restrepo A., Queiroz-Telles F., Nucci M. Epidemiology of endemic systemic fungal infections in Latin America. Med. Mycol. 2011; 49(8):785—98. DOI: 10.3109/13693786.2011.577821.

4. Gopalakrishnan R., Nambi P.S., Ramasubramanian V, Ghafur A., Parameswaran A. Histoplasmosis in India: truly uncommon or uncommonly recognized? J. Assoc. Physicians India. 2012; 60:25—8. PMID: 23777021.

5. Pan B Chen M., Pan W., Liao W. Histoplasmosis: a new endemic fungal infection in China? Review and analysis of cases. Mycoses. 2013; 56(3):212—21. DOI: 10.1111/myc.12029.

6. Norkaew T., Ohno H., Sriburee P., Tanabe K., Tharavichitkul P., Takarn P., Puengchan T., Bumrungsri S., Miyazaki Y. Detection of environmental sources of Histoplasma capsulatum in Chiang Mai, Thailand, by nested PCR. Mycoathologia. 2013; 176(5—6):395—402. DOI: 10.1007/s11046-013-9701-9.

7. Wang Y, Pan B., Wu J., Bi X., Liao W., Pan W., Gu J. Detection and phylogenetic characterization of a case of Histoplasma capsulatum Infection in mainland China. Am. J. Trop. Med. Hyg. 2014; 90(6):1180—3. DOI: 10.4269/ajtmh.14-0048.

8. De D., Nath U.K. Disseminated histoplasmosis in immunocompetent individuals — not a so rare entity, in India. Mediterr. J. Hematol. Infect. Dis. 2015; 7(1):e2015028: DOI: 10.4084/MJHID.2015.028.

9. Jung E.J., Park D.W., Choi J.W., Choi W.S. Chronic cavitary pulmonary histoplasmosis in a non-HIV and immunocompromised patient without overseas travel history. Yonsei Med. J. 2015; 56(3):871—4. DOI: 10.3349/ymj.2015.56.3.871.

10. Lofgren S.M., Kirsch E.J., Maro V.P., Morrissey A.B., Msuya L.J., Kinabo G.D., Saganda W., Diefenthal H.C., Ramadhani H.O., Wheat L.J., Crump J.A. Histoplasmosis among hospitalized febrile patients in northern Tanzania. Trans. R. Soc. Trop. Med. Hyg. 2012; 106(8):504—7. DOI: 10.1016/j.trstmh.2012.05.009.

11. Cottle L.E., Gkrania-Klotsas E., Williams H.J., Brindle H.E., Carmichael A.J., Fry G., Beeching N.J. A multinational outbreak of histoplasmosis following a biology field trip in the Ugandan rainforest. J. Travel. Med. 2013; 20(2):83—7. DOI: 10.1111/jtm.12012.

12. Ashbee H.R., Evans E.G., Viviani M.A., Dupont B., Chryssanthou E., Surmont I., Tomsikova A., Vachkov P., Enero B., Zala J., Tintelnot K., The ECMM Working Group on Histoplasmosis. Histoplasmosis in Europe: report on an epidemiological survey from the European Confederation of Medical Mycology Working Group. Med. Mycol. 2008; 46(1):57—65. DOI: 10.1080/13693780701591481.

13. Rodriguez-Tudela J.L., Alastruey-Izquierdo A., Gago S., Cuenca-Estrella M., Leon C., Miro J.M., Nunez Boluda A., Ruiz Camps I., Sole A., Denning D.W., The University of Manchester in association with the LIFE program. Burden of serious fungal infection in Spain. Clin. Microbiol. Infect. 2015; 21(2):183—9. DOI: 10.1016/j.cmi.2014.07.013.

14. Damasceno L.S., Leitao T.M., Taylor M.L., Muniz M.M., Zancope-Oliveira R.M. The use of genetic markers in the molecular epidemiology of histoplasmosis: a systematic review. Eur. J. Clin. Microbiol. Infect. Dis. 2016; 35(1):19—27. DOI: 10.1007/s10096-015-2508-5.

15. Knox K.S., Hage C.A. Histoplasmosis. Proc. Am. Thorac. Soc. 2010; 7(3):169—72. DOI: 10.1513/pats.200907-069AL.

16. Horwath M.C., Fecher R.A., Deepe G.S. Jr. Histoplasma capsulatum, lung infection and immunity. Future Microbiol. 2015; 10(6):967—75. DOI: 10.2217/fmb.15.25.

17. de M. Teixeira M., Patane J.S., Taylor M.L., Gomez B.L., Theodoro R.C., de Hoog S., Engelthaler D.M., Zancope-Oliveira R.M., Felipe M.S., Barker B.M. Worldwide phylogenetic distributions and population dynamics of the genus Histoplasma. PLoS Negl. Trop. Dis. 2016; 10(6):e0004732. DOI: 10.1371/journal.pntd.0004732.

18. Vite-Garin T., Estrada-Barcenas D.A., Cifuentes J., Taylor M.L. The importance of molecular analysis for understanding the genetic diversity ofHistoplasma capsulatum: an overview. Rev. Iberoam. Micol. 2014; 31(1):11—5. DOI: 10.1016/j.riam.2013.09.013.

19. Taylor M.L., Hernandez-Garcia L., Estrada-Barcenas D., Salas-Lizana R., Zancope-Oliveira R.M., de la Cruz G.S., Galvao-Dias M.A., Curiel-Quesada E., Canteros C.E., Bojorquez-Torres G., Bogard-Fuentes C.A., Zamora-Tehozol E. Genetic diversity of Histoplasma capsulatum isolated from infected bats randomly cap-tured in Mexico, Brazil and Argentina, using the polymorphism of (GA) microsatellite and its flanking regions. Fungal Biol. 2012; 116(2):308—17. DOI: 10.1016/j.funbio.2011.12.004.

20. Arunmozhi Balajee S., Hurst S.F., Chang L.S., Miles M., Beeler E., Hale C., Kasuga T., Benedict K., Chiller T., Lindsley M.D. Multilocus sequence typing of Histoplasma capsulatum in formalin-fixed paraffin-embedded tissues from cats living in non-endemic regions reveals a new phylogenetic clade. Med. Mycol. 2013; 51(4):345—51. DOI: 10.3109/13693786.2012.733430.

21. Eisenberg T., Seeger H., Kasuga T., Eskens U., Sauerwald C., Kaim U. Detection and characterization of Histoplasma capsulatum in a German badger (Meles meles) by ITS-sequencing and multilocus sequencing analysis. Med Mycol. 2013; 51(4):337—44. DOI: 10.3109/13693786.2012.723831.

22. Hage C.A., Ribes J.A., Wengenack N.L., Baddour L.M., Assi M., McKinsey D.S., Hammoud K., Alapat D., Babady N.E., Parker M., Fuller D., Noor A., Davis T.E., Rodgers M., Connolly P.A., El Haddad B., Wheat L.J. A multicenter evaluation of tests for diagnosis of histoplasmosis. Clin. Infect. Dis. 2011; 53(5):448—54. DOI: 10.1093/cid/cir435.

23. Hage C.A., Davis T.E., Fuller D., Egan L., Witt J.R. 3rd, Wheat L.J., Knox K.S. Diagnosis of histoplasmosis by antigen detection in BAL fluid. Chest. 2010; 137(3):623—8. DOI: 10.1378/chest.09-1702.

24. Durkin M., Connolly P., Kuberski T., Myers R., Kubak B.M., Bruckner D., Pegues D., Wheat L.J. Diagnosis of coccidioidomycosis with use of the Coccidioides antigen enzyme immunoassay. Clin. Infect. Dis. 2008; 47(8):e69-73. DOI: 10.1086/592073.

25. Frias-De-Leon M.G., Ramtiez-Barcenas J.A., Rodriguez-Arellanes G., Velasco-Castrejon O., Taylor M.L., Reyes-Montes M.D.R. Usefulness of molecular markers in the diagnosis of occupational and recreational histoplasmosis outbreaks. Folia Microbiol. (Praha). 2017; 62(2):111-6. DOI: 10.1007/s12223-016-0477-4.

26. Assi M., Martin S., Wheat L.J., Hage C., Freifeld A., Avery R., Baddley J.W., Vergidis P., Miller R., Andes D., Young J.A., Hammoud K., Huprikar S., McKinsey D., Myint T., Garcia-Diaz J., Esguerra E., Kwak E.J., Morris M., Mullane K.M., Prakash V, Burdette S.D., Sandid M., Dickter J., Ostrander D., Antoun S.A., Kaul D.R. Histoplasmosis after solid organ transplant. Clin. Infect. Dis. 2013; 57(11):1542—9. DOI: 10.10937cid/cit593.

27. Sampaio Ide L., Freire A.K., Ogusko M.M., Salem J.I., De Souza J.V Selection and optimization of PCR-based methods for the detection of Histoplasma capsulatum var. capsulatum. Rev. Iberoam. Micol. 2012; 29(T):34-9. DOI: 10.1016/j.riam.2011.03.008.

28. Frias De Leon M.G., Arenas Lopez G., Taylor M.L., Acosta Altamirano G., Reyes-Montes Mdel R. Development of specific sequence-characterized amplified region markers for detecting Histoplasma capsulatum in clinical and environmental samples. J. Clin. Microbiol. 2012; 50(3):673-9. DOI: 10.1128/JCM.05271-11.

29. Dantas K.C., Freitas R.S., Moreira A.P, Silva M.V., Benard G., Vasconcellos C., Criado P.R. The use of nested Polymerase Chain Reaction (nested PCR) for the early diagnosis of Histoplasma capsulatum infection in serum and whole blood of HIV-positive patients. An. Bras. Dermatol. 2013; 88(1):141-3. DOI: 10.1590/s0365-05962013000100025.

30. Gago S., Esteban C., Valero C., Zaragoza O., Puig de la Bellacasa J., BuitTago M.J. A multiplex real-time PCR for identification of Pneumocystis jirovecii, Histoplasma capsulatum and Cryptococcus neoformans/Cryptococcus gattii in samples from AIDS patients with opportunistic pneumonia. J. Clin. Microbiol. 2014; 52(4):1168-76. DOI: 10.1128/JCM.02895-13.

31. Munoz C., Gomez B.L., Tobon A., Arango K., Restrepo A., Correa M.M., Muskus C., Cano L.E., Gonzalez A. Validation and clinical application of a molecular method for identification of Histoplasma capsulatum in human specimens in Colombia, South America. Clin. Vaccine. Immunol. 2010; 17(1):62-7. DOI: 10.1128/CVI.00332-09.

32. Hernandez J.M., Munoz-Cadavid C.O., Hernandez D.L., Montoya C., Gonzalez A. Detection of Histoplasma cap-sulatum DNA in peripheral blood from a patient with ocular histoplasmosis syndrome. Med. Mycol. 2012; 50(2):202-6. DOI: 1(13109/13693786.2011.593050.

33. Koepsell S.A., Hinrichs S.H., Iwen P.C. Applying a real-time PCR assay for Histoplasma capsulatum to clinically relevant formalin-fixed paraffin-embedded human tissue. J. Clin. Microbiol. 2012; 50(10):3395-7. DOI: 10.1128/JCM.01705-12.

34. Scheel C.M., Zhou Y, Theodoro R.C., Abrams B., Balajee S.A., Litvintseva A.P. Development of a loop-mediated isothermal amplification method for detection of Histoplasma capsulatum DNA in clinical samples. J. Clin. Microbiol. 2014; 52(2):483-8. DOI: 10.1128/JCM.02739-13.

35. Furuie J.L., Sun J., do Nascimento M.M., Gomes R.R., Waculicz-Andrade C.E., Sessegolo G.C., Rodrigues A.M., Galvao-Dias M.A., de Camargo Z.P., Queiroz-Telles F., Najafzadeh M.J., de Hoog S.G., Vicente V.A. Molecular identification of Histoplasma capsulatum using rolling circle amplification. Mycoses. 2016; 59(1):12-9. DOI: 10.1111/myc.12426.

36. Crockett D.K., Kushnir M.M., Cloud J.L., Ashwood E.R., Rockwood A.L. Identification of histoplasma-specific pep¬tides in human urine. Int. J. Pept. 2012; 2012:621329. DOI: 10.1155/2012/621329.

37. Buitrago M.J., Canteros C.E., Frias De Leon G., Gonzalez A., Marques-Evangelista De Oliveira M., Munoz C.O., Ramirez J.A., Toranzo A.L., Zancope-Oliveira R., Cuenca-Estrella M. Comparison of PCR protocols for detecting Histoplasma capsulatum DNA through a multicenter study. Rev. Iberoam. Micol. 2013; 30(4):256-60. DOI: 10.1016/j.riam.2013.03.004.

38. Efias N.A., Cuestas M.L., Sandoval M., Poblete G., Lopez-Daneri G., Jewtuchowicz V, Iovannitti C., Mujica M.T. Rapid iden-tification of Histoplasma capsulatum directly from cultures by multi-plex PCR. Mycopathologia. 2012; 174(5-6):451-56. DOI: 10.1007/s11046-012-9567-2.

39. Simon S., Veron V., Boukhari R., Blanchet D., Aznar C. Detection of Histoplasma capsulatum DNA in human samples by real-time polymerase chain reaction. Diagn. Microbiol. Infect. Dis. 2010; 66(3): 268-73. DOI: 10.1016/j.diagmicrobio.2009.10.010.

40. Holbrook E.D., Edwards J.A., Youseff B.H., Rappleye C.A. Definition of the extracellular proteome of pathogenic-phase Histoplasma capsulatum. J. Proteome Res. 2011; 10(4):1929-43. DOI: 10.1021/pr1011697.

41. Ohno H., Tanabe K., Umeyama T., Kaneko Y, Yamagoe S, Miyazaki Y. Application of nested PCR for diagnosis of histoplas-mosis. J. Infect. Chemother. 2013; 19(5):999-1003. DOI: 10.1007/s10156-013-0548-2.

42. Muraosa Y, Toyotome T., Yahiro M., Watanabe A., Shikanai-Yasuda M.A., Kamei K. Detection of Histoplasma capsulatum from clinical specimens by cycling probe-based real-time PCR and nested real-time PCR. Med. Mycol. 2016; 54(4):433-8. DOI: 10.1093/mmy/myv106.

43. Muniz Mde M., Morais E. Silva Tavares P., Meyer W., Nosanchuk J.D., Zancope-Oliveira R.M. Comparison of different DNA-based methods for molecular typing of Histoplasma capsulatum. Appl. Environ. Microbiol. 2010; 7б(13):4438-4/. DOI: 10.1128/AEM.02004-09.

44. Munoz B., Martinez M.A., Palma G., Ramtiez A., Frias M.G., Reyes M.R., Taylor M.L., Higuera A.L., Corcho A., Manjarrez M.E. Molecular characterization of Histoplasma capsulatum isolated from an outbreak in treasure hunters. BMC Infect. Dis. 2010; 10:264. DOI: 10.1186/1471-2334-10-264.

45. Reyes-Montes M.R., Rodriguez-Arellanes G., Perez-Torres A., Rosas-Rosas A.G., Paras-Garda A., Juan-Salles C., Taylor M.L. Identification of the source of histoplasmosis infection in two captive maras (Dolichotis patagonum) from the same colony by using molecular and immunologic assays. Rev. Argent. Microbiol. 2009; 41(2):102-4. PMID: 19623900.

46. Balajee S.A., Borman A.M., Brandt M.E., Cano J., Cuenca-Estrella M., Dannaoui E., Guarro J., Haase G., Kibbler C.C., Meyer W., O’Donnell K., Petti C.A., Rodriguez-Tudela J.L., Sutton D., Velegraki A., Wickes B.L. Sequence-based identification of Aspergillus, Fusarium, and Mucorales species in the clinical mycolosy laboratory: where are we and where should we go from here? J. Clin. Microbiol. 2009; 47(4):877-84. DOI: 10.1128JCM.01685-08.