Evaluating and optimizing Acid-pH and Direct Lysis RNA extraction for SARS-CoV-2 RNA detection in whole saliva.

COVID-19 has been a global public health and economic challenge. Screening for the SARS-CoV-2 virus has been a key part of disease mitigation while the world continues to move forward, and lessons learned will benefit disease detection beyond COVID-19. Saliva specimen collection offers a less invasive, time- and cost-effective alternative to standard nasopharyngeal swabs. We optimized two different methods of saliva sample processing for RT-qPCR testing. Two methods were optimized to provide two cost-efficient ways to do testing for a minimum of four samples by pooling in a 2.0 mL tube and decrease the need for more highly trained personnel. Acid-pH-based RNA extraction method can be done without the need for expensive kits. Direct Lysis is a quick one-step reaction that can be applied quickly. Our optimized Acid-pH and Direct Lysis protocols are reliable and reproducible, detecting the beta-2 microglobulin (B2M) mRNA in saliva as an internal control from 97 to 96.7% of samples, respectively. The cycle threshold (Ct) values for B2M were significantly higher in the Direct Lysis protocol than in the Acid-pH protocol. The limit of detection for N1 gene was higher in Direct Lysis at ≤ 5 copies/µL than Acid-pH. Saliva samples collected over the course of several days from two COVID-positive individuals demonstrated Ct values for N1 that were consistently higher from Direct Lysis compared to Acid-pH. Collectively, this work supports that each of these techniques can be used to screen for SARS-CoV-2 in saliva for a cost-effective screening platform.

SNHG7 and FAIM2 are up-regulated and co-expressed in colorectal adenocarcinoma tissues.

BACKGROUND: The global incidence of colorectal cancer (CRC) is expected to be increased by 60% until a few years. Despite the advances in surgical and chemotherapy techniques, a significant proportion of patients with CRC have poor responses to treatments. These are the reasons that prove the importance of identifying molecular bio-markers as potential therapeutic targets. Long non-coding RNAs (lnc RNAs) participate in the initiation, development, progression, and metastasis of cancers such as CRC. Hence, this class of noncoding RNAs is known as bio-marker for cancer dia-gnosis and prognosis. MATERIALS AND METHODS: In this experimental study, the extraction of total RNA from tissues, synthesis of complementary DNA as well as quantitative real-time polymerase chain reaction (qRT- PCR) were performed. Comparative cycle threshold method was applied to quantify the expression level of lncRNA-SNHG7 and FAIM2. The relative amount of lncRNA-SNHG7 and FAIM2 was calculated using the equation 2 -DDCT. RESULTS: In this study, by qRT PCR, we concluded that the expression level of SNHG7, as a recently identified lncRNA and FAIM2 were increased in colorectal cancer tissues compared with normal adjacent tissues. CONCLUSION: Our study indicates the potential importance of SNHG7 and FAIM2 expression for more studies in future.