ABSTRACT
BackgroundThe current increase in the spread of (SARS-CoV-2) critically needs a multitarget diagnostic assays to promote analytical sensitivity to facilitate the public health actions. ObjectiveThe aim of this study was to develop a new primer-probe set targeting N gene of SARS-CoV-2 to improve the sensitivity for detection of COVID-19(Corona Virus Disease 2019)in multiplex rRT-PCR (Reversetranscript Realtime PCR) and ddPCR (Droplet Digital PCR). ResultsWe designed primers/probes set N(LZU3) targeting the N gene of 2019-nCov and proved its sensitivity in both rRT-PCR and ddPCR. When the quantity of template was 105 copies/reaction, the mean Ct value of N(LZU3) was 32.563, the detection rate was 91.7%. If the quantity of template was 52.5 copies/reaction, the mean Ct value of N(LZU3) was 33.835, and the detection rate was 83.3%, which were similar with that of N(CDC) and N(USA). The calculated lower limit of detection (LOD) of the new primer-probe set N(LZU3) used in rRT-PCR was 118 copies/reaction. We also did one-step ddPCR for detection the same serial dilution of RNA template. It shows good linearity for primer/probe sets N(LZU3). The calculated lower limit of detection (LOD) of N(LZU3) was 22.4 copies/reaction, which was 1.12 copies/ul. ConclusionThe novel primer-probe set(LZU3) targeting N gene of SARS-CoV-2 could be both used in rRT-PCR and ddPCR with better sensitivity, furthermore, ddPCR method had higer sensitivity than rRT-PCR, hence it could significantly improve SARS-CoV-2 detection efficiency in low virus load and asymptomatic infection.
ABSTRACT
Objective To investigate the inhibitory effects of trichostatin A(TSA)on bladder cancer cell lines and its synergetic effect with anticancer drugs in treating bladder cancer in vitro and in vivo.Methods The inhibitory effects of TSA on human bladder cancer cell lines in vitro were detected by MTT assay.Hoechst staining was used to observe morphology for apoptotic cells after TSA treatment.Western blot was used to detect expression of acetyl-histone H3 and survivin.In vivo synergetic effects of TSA with anticancer drugs were detected in bladder cancer model rats.Results TSA significantly inhibited growth of bladder cancer cell lines in concentration and time dependent manner.Better results of tumor inhibition have been achieved when it was combined with DDP,MMC and ADM than used alone.After TSA treatment,the survivin expression in bladder cancer cells decreased and acetyl-histone H3 expression increased.Intravesical application of TSA combined with MMC can significantly inhibited tumor growth and progression.Conclusion TSA has direct anti-cancer effect and can enhance the action of several chemotherapy agents markedly.TSA may be an excellent candidate agent for intravesical application to treat bladder cancer.
