Design and optimization of an improved qPCR assay for the detection of Histoplasma capsulatum-Reference-Cited by-同舟云学术

Design and optimization of an improved qPCR assay for the detection of Histoplasma capsulatum

Published:2023-10-13 Issue:2 Volume:13 Page:37-44
ISSN:2062-509X
Container-title:European Journal of Microbiology and Immunology
language:
Short-container-title:EuJMI

Author:

da Silva Bianca Kelly Neves Izidro¹,Alves Cortez Ana Cláudia²,Oliveira Luciana Aires²,de Almeida Juan Diego Ribeiro²,de Souza Érica Simplício³,Cruz Katia Santana⁴,Jackisch Matsuura Ani Beatriz⁵,Ogusku Mauricio Morishi⁶,Solórzano Chavarría Karolina Jeaneth⁷⁸,Baía-da-Silva Djane¹⁷⁸⁹¹⁰,Bassat Quique¹¹¹²¹³¹⁴¹⁵,Guimarães Lacerda Marcus Vinícius⁷⁸⁹¹⁶,Frickmann Hagen¹⁷¹⁸,de Souza João Vicente Braga²^ORCID

Affiliation:

1. Graduate Program in Pharmaceutical Sciences, Universidade Federal do Amazonas-UFAM, Manaus, Brazil

2. Mycology Laboratory, Instituto Nacional de Pesquisas da Amazonia–INPA, Manaus, Brazil

3. Superior School of Technology, Universidade do Estado do Amazonas -UEA, Manaus, Brazil

4. Medical Mycology Laboratory, Fundação de Medicina Tropical Doutor Heitor Vieira Dourado-FMT-HVD, Manaus, Brazil

5. Microbial Diversity Laboratory of the Amazon with Health Importance, Leônidas and Maria Deane Institute-Fiocruz-AM, Manaus, Amazonas, Brazil

6. Mycobacteriology Laboratory, Instituto Nacional de Pesquisas da Amazonia–INPA, Manaus, Brazil

7. Graduate Program in Tropical Medicine, Fundação de Medicina Tropical Doutor Heitor Vieira Dourado-FMT-HVD, Manaus, Brazil

8. Carlos Borborema Clinical Research Unit, Fundação de Medicina Tropical Doutor Heitor Vieira Dourado-FMT-HVD, Manaus, Brazil

9. Instituto Leônidas Maria Deane- ILMD, Fiocruz-AM, Manaus, Brazil

10. Pharmacy Faculty, Universidade Nilton Lins, Manaus, Brazil

11. ISGlobal, Hospital Clínic – Universitat de Barcelona, Barcelona, Spain

12. Centro de Investigação em Saúde de Manhiça (CISM), Maputo, Mozambique

13. ICREA, Pg. Lluís Companys 23, 080100 Barcelona, Spain

14. Pediatrics Department, Hospital Sant Joan de Déu, Universitat de Barcelona, Esplugues, Barcelona, Spain

15. Consorcio de Investigación Biomédica en Red de Epidemiología y Salud Pública (CIBERESP), Madrid, Spain

16. University of Texas Medical Branch, Galveston, USA

17. Institute for Medical Microbiology, Virology and Hygiene, University Medicine Rostock, Rostock, Germany

18. Department of Microbiology and Hospital Hygiene, Bundeswehr Hospital, Hamburg, Germany

Abstract

AbstractBackgroundThis study aimed at improving a real-time polymerase-chain-reaction (qPCR) assay for the detection of Histoplasma capsulatum, a fungal pathogen that can cause severe respiratory infections in humans, in clinical and soil samples.MethodsPrimer and probes were in-silico designed, in-silico and in-vitro evaluated including clinical biopsy materials and finally subjected to a real-world application with collected soil samples.ResultsApplying the qPCR assay with liver and lung biopsies from 71 patients each, including 59 patients infected with human immunodeficiency virus (HIV), as well as with Sabouraud (SAB) agar culture as the diagnostic reference standard, diagnostic accuracy of the qPCR assay of 100% (5/5) sensitivity and 96% (63/66) specificity for liver samples and 100% (4/4) sensitivity and 94% (63/67) specificity for the lung samples was recorded. When applying the assay with soil samples from caves near of Presidente Figueiredo city, Amazonas, Brazil, one sample from the Maroaga cave was confirmed as positive.ConclusionsThe improved qPCR assessed in this study was successful in detecting H. capsulatum with high efficiency and accuracy in in-vitro evaluation, including the identification of the target pathogen in both clinical and environmental samples.

Funder

CNPQ

Fundação de Amparo a Pesquisa do Amazonas

Publisher

Akademiai Kiado Zrt.

Subject

Applied Mathematics,General Mathematics

Link

https://akjournals.com/downloadpdf/journals/1886/13/2/article-p37.xml

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