Implementation of in-house Methods for Isolating Fungal DNA of Clinical Samples

David Koffi^1, , Yayé Yapi Guillaume², Francis K. Kouadjo^{1, 2}, Koui S Tossea¹, Andre O. Toure¹ and Allico J Djaman²

¹Parasitology and Mycology Department, Institut Pasteur, Abidjan, Côte d’Ivoire

²Agrovalorisation Laboratory, University Jean Lorougnon Guédé, Daloa, Cote d’Ivoire

Cite this paper:
David Koffi, Yayé Yapi Guillaume, Francis K. Kouadjo, Koui S Tossea, Andre O. Toure and Allico J Djaman. Implementation of in-house Methods for Isolating Fungal DNA of Clinical Samples. American Journal of Biomedical Research. 2023; 11(1):7-13. doi: 10.12691/ajbr-11-1-2

Abstract

Over 200 species of fungi are responsible for a variety of infections that can occur in various parts of the human body. There are several phenotypic methods for identifying these fungal elements; however, these approaches have limitations. Molecular methods are now routinely used in well-equipped mycology laboratories. However, the first step in isolating genetic material can often be costly, can suffer from external DNA contamination and some components have toxicity for personnel. The general objective of this work was to identify the best local method for isolating genetic material from fungi based on cost, yield and time-consuming criteria. A total of ten (10) different nucleic acid isolation methods were tested. Those tests using thermal or mechanical shock for cell lysis delivered better quality than those using chemical lysis. Thus, based on our criteria, the best methods for nucleic acid isolation and purification of fungal elements were cetyltrimethylammonium bromide (CTAB) combined with sterilized sea sand (less expensive) and the chelator Chelex® coupled with glass beads (faster).

Keywords:
thermal shock mechanical shock fungal species DNA extraction

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