Abstract
Good quality RNA needs to be obtained in order to study gene expression. Different RNA extraction methods have been described, but RNA quality and yield may vary among the different techniques and biological study species. To date, there is no standardized method for extraction and purification of RNA from Candida genus yeasts. The few available papers on the subject apply mainly to filamentous fungi and have produced poor results for extraction techniques based on manual or in-house IVD methods. The aim of this study was therefore to compare two commercial RNA extraction and purification systems using silica columns (Qiagen and Zymo Research) with Candida parapsilosis sensu stricto as model organism. This yeast has been identified in recent papers as the second most frequently isolated Candida species in the oral cavity. In the past decade, it has been the object of increasing medical interest because it is one of the main causes of candidemia in both adults and preterm neonates. In view of this background, we consider the study of Candida parapsilosis sensu stricto transcriptome and its variations according to environmental changes to be a priority. In this experimental study, 19 fungal isolates were processed using Qiagen and 17 isolates using Zymo Research. The results suggest that Qiagen lysis buffer RLT is essential for obtaining better quality RNA product.
Highlights
Developments in molecular biology have enabled molecular techniques to be used in mycological studies, thereby promoting more precise diagnoses in shorter times and above all, enabling culturable fungi with low abundance as well as non-culturable fungi to be identified through analysis of their genetic material
RNA extraction is critical for fungal cells because their cell wall characteristics differ according to genus, and the processes must be optimized for each particular case
RRNA band intensity bought to be even for all samples regardless of the commercial system used, since the expression of that region should be consistent among all strains, as they all belong to the same species. These results suggest that the RNA obtained using the Zymo Research protocol may have reduced the efficiency of the RT-PCR reaction
Summary
Developments in molecular biology have enabled molecular techniques to be used in mycological studies, thereby promoting more precise diagnoses in shorter times and above all, enabling culturable fungi with low abundance as well as non-culturable fungi to be identified through analysis of their genetic material. Techniques for analysis of gene expression have enabled the study of gene function, providing understanding of interactions between the host and its mycobiome, and the response of each fungal species to different environmental conditions. Good quality RNA is needed to study gene expression. It is important to use the best possible RNA extraction method, since any contaminants such as RNases, proteins, polysaccharides and genomic DNA may affect RNA quality and reduce the efficiency of its amplification. RNA extraction is critical for fungal cells because their cell wall characteristics differ according to genus, and the processes must be optimized for each particular case. RNA is highly labile and less stable than DNA [1]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
