Abstract
The fungus Paracoccidioides brasiliensis is the pathogen of paracoccidioidomycosis (PCM), a systemic mycosis prevalent in Latin America. The loop‐mediated isothermal amplification method (LAMP) was used in this study to detect the presence of P. brasiliensis in sputa samples from patients with chronic PCM, suspected PCM, and a negative control. The target P. brasiliensis gp43 gene was amplified in less than 4 hr in 11 of 18 sputa samples tested. The LAMPmethod had the advantage of speed and simplicity compared with the classic diagnostic methods such as the histopathological test or biological material culture and did not require sophisticated technical apparatus. It would be an important aid in cases where immediate treatment would mean patient survival, especially in immune‐suppressed patients. J. Clin. Lab. Anal. 23:139–143 2009. © 2009 Wiley‐Liss, Inc.
Keywords: gp43 gene, Paracoccidioides brasiliensis, paracoccidiomycosis, LAMP
REFERENCES
- 1. Restrepo A, McEwen JG, Castaneda E. The habitat of Paracoccidioides brasiliensis: How far from solving the riddle? Med Mycol 2001;39:233–241. [DOI] [PubMed] [Google Scholar]
- 2. Guezelle E. Aislamiento de Paracoccidioides sp. de heces de pinguino de la Antártida, res. B2. In: Resúmenes IV Encuentro Internacional sobre Paracoccidioidomicosis. Caracas: Venezuela, 1989.
- 3. Terçarioli GR, Bagagli E, Reis GM, et al. Ecological study of Paracoccidioides brasiliensis in soil: Growth ability, conidia production and molecular detection. BMC Microbiol 2007;7 :92. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. Ferreira MS, Freitas LH, Lacaz CS, et al. Isolation and characterization of a Paracoccidioides brasiliensis strain from a dog food probably contaminated with soil in Uberlandia, Brazil. J Med Vet Mycol 1990;28:253–256. [DOI] [PubMed] [Google Scholar]
- 5. Grosse E, Tamsitt J. Paracoccidioides brasiliensis recovered from the intestinal tract of three bats (Artibeus lituratus) in Colombia S.A. Sabouraudia 1965;4:124–125. [DOI] [PubMed] [Google Scholar]
- 6. Bagagli E, Sano A, Coelho KIR, et al. Isolation of Paracoccidioides brasiliensis from armadillos (Dasypus novemcinctus) in an endemic area of paracoccidioidomycosis. Am J Trop Med Hyg 1998;58:505–512. [DOI] [PubMed] [Google Scholar]
- 7. Calle D, Rosero DS, Orozco LC, Camargo D, Castaneda E, Restrepo A. Paracoccidioidomycosis in Colombia: An ecological study. Epidemiol Infect 2001;126:309–315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8. Coutinho ZF, Silva D, Lazera M, Petri V, Oliveira RM, Sabroza PCl. Paracoccidioidomycosis mortality in Brazil (1980–1995). Cad Saúde Pública 2002;18:1441–1454. [DOI] [PubMed] [Google Scholar]
- 9. Londero AT, Del Negro G. Paracoccidioidomicose. J Pneumol 1986;12:41–60. [Google Scholar]
- 10. Franco M. Host‐parasite relationships in paracoccidioidomycosis. J Med Vet Mycol 1986;25:5–18. [DOI] [PubMed] [Google Scholar]
- 11. Elias Costa MR, Da Silva Lacaz C, Kawasaki M, De Camargo ZP. Conventional versus molecular diagnostic tests. Med Mycol 2000;38:139–145. [PubMed] [Google Scholar]
- 12. Wanke B, Londero AT. Paracoccidioides brasiliensis In: Ajello L, Hay RJ, editors. Topley & Wilson's Microbiology and Microbial Infections, Medical Mycology, Vol. 4, Ninth edition London, England: Arnold, 1998. [Google Scholar]
- 13. Ajello L, Polonelli L. Imported paracoccidioidomycosis: A public health problem in non‐endemic areas. Eur J Epidemiol 1985;1:160–165. [DOI] [PubMed] [Google Scholar]
- 14. Endo S, Komori T, Ricci G, et al. Detection of gp43 of Paracoccidioides brasiliensis by the loop‐mediated isothermal amplification (LAMP) method. FEMS Microb Lett 2004;23:93–97. [DOI] [PubMed] [Google Scholar]
- 15. Igarashi T, Kurose T, Itabashi K, et al. A case of chronic pulmonary paracoccidioidomycosis. Nihon Kokyuki Gakkai Zasshi 2004;42:629–633. [PubMed] [Google Scholar]
- 16. Camargo ZP, Franco MF. Current knowledge on pathogenesis and immunodiagnosis of paracoccidioidomycosis. Rev Iberam Micol 2000;17:41–48. [PubMed] [Google Scholar]
- 17. Brummer E, Castaneda E, Restrepo A. Paracoccidioidomycosis: An update. Clin Microbiol Rev 1993;6:89–117. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18. Siqueira AM. Avaliação da sensibilidade e especificidade de algumas provas sorológicas no diagnóstico, prognóstico e controle de cura da paracoccidioidomicose. Caracterização do antígeno E2 de Paracoccidioides brasiliensis [Tese de Doutorado]. São Paulo: Universidade de São Paulo, 1982.
- 19. Neves AR, Mamoni RL, Rossi CL, Camargo ZP, Blotta MHS. Negative immunodiffusion test results obtained with sera of paracoccidioidomycosis patients may be related to low‐affinity immunoglobulin G2 antibodies directed against carbohydrate epitopes. Clin Diagn Lab Immunol 2003;10:802–807. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20. Silva SHM, de Mattos Grosso D, Lopes JD, et al. Detection of Paracoccidioides brasiliensis gp70 circulating antigen and follow‐up of patients undergoing antimycotic therapy. J Clin Microbiol 2004;42:4480–4486. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21. Gomes GM. Padronização da técnica de PCR para diagnostico de paracoccidioidomicose baseada em sequências do gene da gp43 de Paracoccidioides brasiliensis [Dissertação de mestrado] São Paulo: Escola Paulista de Medicina, 1999.