| dc.description.abstract | Saliva is one of the prospective samples used to identify respiratory infections
and diagnose human diseases, including diabetes, cardiovascular disease, and
cancer. Saliva has a significant benefit over various other samples, such as
serum and nasopharyngeal swab (NPS), because it is collected quickly and
painlessly. In addition, we require a rapid and accurate diagnostic test for viral
detection to respond and prevent infectious diseases immediately before
spreading. Therefore, our lab develops a direct one-step RT-qPCR from saliva
samples, which is a rapid and cost-effective test that might be utilized in the
case of a future pandemic. Nevertheless, the significant factor in RT-qPCR is the
internal control gene, which prevents false negative results and ensures a
reliable assay result. Typically, housekeeping genes (HKG) were used as ICGs
because their expression was assumed to be stable across various tissue types,
ages, and therapies. However, housekeeping genes’ expression stability varies
depending on cell type, biological conditions, and experimental conditions.
Researchers did not validate housekeeping genes as the internal control for
direct RT-qPCR from saliva samples; consequently, it is critical to verify a stable
gene as the internal control gene for direct one-step RT-qPCR. In this study, we
discovered that RNase P is the stable gene that might serve as our internal
control for the assay. Unfortunately, the RNase P primer from the CDC can bind
to DNA; therefore, it is inappropriate to use it in direct one-step RT-qPCR from
whole saliva. Furthermore, our group designs and proves novel primers for the
RNase P gene, which targets RNA exclusively. Future development of the direct
one-step RT-qPCR assay for infectious disease detection will be facilitated by the
evaluation and validation of the internal control gene RNase P. | en_US |
