Comparison of diagnostic methods to detect Histoplasma capsulatum in serum and blood samples from AIDS patients

PLoS One. 2018 Jan 17;13(1):e0190408. doi: 10.1371/journal.pone.0190408. eCollection 2018.

Abstract

Background: Although early and rapid detection of histoplasmosis is essential to prevent morbidity and mortality, few diagnostic tools are available in resource-limited areas, especially where it is endemic and HIV/AIDS is also epidemic. Thus, we compared conventional and molecular methods to detect Histoplasma capsulatum in sera and blood from HIV/AIDS patients.

Methodology: We collected a total of 40 samples from control volunteers and patients suspected of histoplasmosis, some of whom were also infected with other pathogens. Samples were then analyzed by mycological, serological, and molecular methods, and stratified as histoplasmostic with (group I) or without AIDS (group II), uninfected (group III), and infected with HIV and other pathogens only (group IV). All patients were receiving treatment for histoplasmosis and other infections at the time of sample collection.

Results: Comparison of conventional methods with nested PCR using primers against H. capsulatum 18S rRNA (HC18S), 5.8S rRNA ITS (HC5.8S-ITS), and a 100 kDa protein (HC100) revealed that sensitivity against sera was highest for PCR with HC5.8S-ITS, followed by immunoblotting, double immunodiffusion, PCR with HC18S, and PCR with HC100. Specificity was equally high for double immunodiffusion, immunoblotting and PCR with HC100, followed for PCR with HC18S and HC5.8-ITS. Against blood, sensitivity was highest for PCR with HC5.8S-ITS, followed by PCR with HC18S, Giemsa staining, and PCR with HC100. Specificity was highest for Giemsa staining and PCR with HC100, followed by PCR with HC18S and HC5.8S-ITS. PCR was less efficient in patients with immunodeficiency due to HIV/AIDS and/or related diseases.

Conclusion: Molecular techniques may detect histoplasmosis even in cases with negative serology and mycology, potentially enabling early diagnosis.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Female
  • HIV Infections / blood*
  • Histoplasma / isolation & purification*
  • Humans
  • Male
  • Middle Aged

Grants and funding

This study was supported by the State of São Paulo Research Foundation (FAPESP) under grant n. 2009/50362-0. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.