Effect of Savirin or Ticagrelor Treatment on the Expression of Commonly Used Reference Genes in

Overview

Journal Microorganisms

Specialty Microbiology

Date 2023 Feb 25

PMID 36838300

Authors

Narayan Pant

Catherine Rush

Jeffrey Warner

Damon P Eisen

Affiliations

Soon will be listed here.

Abstract

Reference genes are frequently used for the normalization of quantitative reverse transcriptase PCR (qRTPCR) data in gene expression studies. is one of the most common causes of biofilm-related infections. Savirin and ticagrelor show in vitro as well as in vivo antibiofilm activity against . The main aim of this study was to identify the most stably expressed reference genes to study the effect of these molecules on genes in a strong biofilm producing isolate isolated from biofilm-related infection. Quantitative real-time PCR was performed by using relative quantification method. Four different algorithms, delta Ct, normfinder, bestkeeper, and genorm, followed by a comprehensive analysis was used to identify the most stable reference genes from a list of sixteen different candidate reference genes. All four algorithms reported different results, with some comparable findings among some methods. In the comprehensive analysis of the results of all the algorithms used, the most stable reference genes found were , , and for savirin treatment experiment and , , and for ticagrelor treatment experiment. The optimal number of reference genes required was two for both the experimental conditions. Despite having some drawbacks, each algorithm can reliably determine an appropriate reference gene independently. However, based on consensus ranking and the required optimal number of reference genes reported, and were the most appropriate reference genes for savirin treatment experiment, and and were most appropriate for ticagrelor treatment experiment. This study provides baseline data on reference genes to study the effect of savirin or ticagrelor treatment on the expression of potential reference genes in . We recommend prior re-validation of reference genes on a case-by-case basis before they can be used.

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