Examining potential confounding factors in gene expression analysis of human saliva and identifying potential housekeeping genes.

ABSTRACT: Isolation of RNA from whole saliva, a non-invasive and easily accessible biofluid that is an attractive alternative to blood for high-throughput biodosimetry of radiological/nuclear victims might be of clinical significance for prediction and diagnosis of disease. In a previous analysis of 12 human samples we identified two challenges to measuring gene expression from total RNA: (1) the fraction of human RNA in whole saliva was low and (2) the bacterial contamination was overwhelming. To overcome these challenges, we performed selective cDNA synthesis for human RNA species only by employing poly(A)+-tail primers followed by qRT-PCR. In the current study, this approach was independently validated on 91 samples from 61 healthy donors. Additionally, we used the ratio of human to bacterial RNA

SUBMITTER: Ostheim P 

PROVIDER: S-EPMC8831573 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature