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. 2016 Feb 24:16:87.
doi: 10.1186/s12879-016-1427-0.

Validation of western blot for Histoplasma capsulatum antibody detection assay

Marcos de Abreu Almeida  1 , Cláudia Vera Pizzini  2 , Lisandra Serra Damasceno  3 , Mauro de Medeiros Muniz  4 , Rodrigo Almeida-Paes  5 , Regina Helena Saramago Peralta  6 , José Mauro Peralta  7 , Raquel de Vasconcelos Carvalhaes Oliveira  8 , Alexandre Gomes Vizzoni  9 , Carla Lourenço Tavares de Andrade  10 , Rosely Maria Zancopé-Oliveira  11
Affiliations

Validation of western blot for Histoplasma capsulatum antibody detection assay

Marcos de Abreu Almeida et al. BMC Infect Dis. .

Abstract

Background: Histoplasmosis is worldwide systemic mycoses caused by the dimorphic fungus Histoplasma capsulatum. The isolation and identification of H. capsulatum in culture is the reference test for histoplasmosis diagnosis confirmation. However, in the absence of it, serology has been used as a presumptive diagnosis through antibody and antigen detection. The purpose of the present study was to validate an immunoassay method (western blot) for antibodies detection in the diagnosis of histoplasmosis.

Methods: To validate the western blot (WB) a study was conducted using 118 serum samples from patients with histoplasmosis and 118 serum controls collected from January 2000 to December 2013 in residents of the Rio de Janeiro State, Brazil. Diagnostic validation parameters were calculated based on the categorization of results obtained in a 2 × 2 table and subjected to statistical analysis. In addition, the viability of deglycosylated histoplasmin antigen (ptHMIN) onto nitrocellulose membranes previously sensitized was evaluated during the same period.

Results: The WB test showed sensitivity of 94.9 %, specificity of 94.1 %, positive predictive value of 94.1 %, negative predictive value of 94.9 %, accuracy of 94.5 %, and almost perfect precision. Besides, the strips have proved to be viable for using at least 5 years after ptHMIN antigen sensitization.

Conclusion: Western blot test using ptHMIN provides sensitive, specific, and faster results. Therefore, could be considered a useful tool in the diagnosis of histoplasmosis being used by public health system, even in situations where laboratory facilities are relatively limited.

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Figures

Fig. 1
Fig. 1
Serum samples tested during validation Western blot immunoassay for diagnosis of histoplasmosis
Fig. 2
Fig. 2
Analysis of the viability of membranes. Lanes MW- Molecular weight; 1- One day; 2- One month; 3- Six months; 4- One year; 5- Two years; 6- Three years; 7- Four years; 8- Five years; 9- Healthy individual (negative control)
See this image and copyright information in PMC

References

    1. Colombo AL, Tobón A, Restrepo A, Queiroz-Telles F, Nucci M. Epidemiology of endemic systemic fungal infections in Latin America. Med Mycol. 2011;49(8):785–798. - PubMed
    1. Gómez B. Histoplasmosis: epidemiology in Latin America. Current Fungal Infect Reports. 2011;5(4):199–205. doi: 10.1007/s12281-011-0073-7. - DOI
    1. Guimarães AJ, Nosanchuk JD, Zancopé-Oliveira RM. Diagnosis of histoplasmosis. Braz J Microbiol. 2006;37(1):1–13. doi: 10.1590/S1517-83822006000100001. - DOI - PMC - PubMed
    1. Damasceno LS, Ramos AN, Jr, Alencar CH, Lima D, Sidrim JJ, Gonçalves M, et al. Disseminated histoplasmosis and aids: relapse and late mortality in endemic area in north-eastern Brazil. Mycoses. 2013;56(5):520–526. doi: 10.1111/myc.12067. - DOI - PubMed
    1. Zancopé-Oliveira RM, Guimarães AJ, Nosanchuk JD, Muniz MM, Albuquerque PC, Almeida-Paes R. Histoplasma. In: Liu D, editor. Molecular detection of human fungal pathogens. Florida: CRC Press; 2011. pp. 236–274.

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