Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assist...Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.展开更多
Objectives To find out the association between the sensorineural hearing loss and coronavirus disease 2019(COVID-19),the expression of ACE2 and TMPRSS2 in hamsters and mice was detected.Design Using the public data fr...Objectives To find out the association between the sensorineural hearing loss and coronavirus disease 2019(COVID-19),the expression of ACE2 and TMPRSS2 in hamsters and mice was detected.Design Using the public data from the National Center for Biotechnology Information and the Global Initiative on Sharing All Influenza Data,the expression of ACE2 and TMPRSS2 at the transcriptomic,DNA,and protein levels of ACE2 in the brain,inner ear,and muscle from the golden Syrian hamsters(Mesocricetus auratus)and mice(Mus musculus)was assessed.Results We identified ACE2 and TMPRSS2 expressed at different levels in the inner ear and brain at DNA and transcriptomic levels of both mice and hamsters.The protein expression from the brain and inner ear showed a similar pattern,while the expression of ACE2 from the inner ear was relatively higher than that from the muscle.Conclusion Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)shows genetic potential to infect the hearing system of rodents and lead to sudden sensorineural hearing loss that can be used as a characteristic to detect asymptomatic patients of COVID-19.展开更多
文摘Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.
基金supported by the Department of Science and Technology of Henan Province(201100312100)Collaborative Innovation Project of Zhengzhou(Zhengzhou University)(18XTZX12004)+1 种基金Scientific Research Project of Epidemic Prevention and Control in Henan(211100310900)Postdoctoral Research Startup Project in Henan Province(202001043).
文摘Objectives To find out the association between the sensorineural hearing loss and coronavirus disease 2019(COVID-19),the expression of ACE2 and TMPRSS2 in hamsters and mice was detected.Design Using the public data from the National Center for Biotechnology Information and the Global Initiative on Sharing All Influenza Data,the expression of ACE2 and TMPRSS2 at the transcriptomic,DNA,and protein levels of ACE2 in the brain,inner ear,and muscle from the golden Syrian hamsters(Mesocricetus auratus)and mice(Mus musculus)was assessed.Results We identified ACE2 and TMPRSS2 expressed at different levels in the inner ear and brain at DNA and transcriptomic levels of both mice and hamsters.The protein expression from the brain and inner ear showed a similar pattern,while the expression of ACE2 from the inner ear was relatively higher than that from the muscle.Conclusion Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)shows genetic potential to infect the hearing system of rodents and lead to sudden sensorineural hearing loss that can be used as a characteristic to detect asymptomatic patients of COVID-19.
