Nucleic acid(DNA and RNA)detection and quantification methods play vital roles in molecular biology.With the development of molecular biology,isothermal amplification of DNA/RNA,as a new molecular biology technology,c...Nucleic acid(DNA and RNA)detection and quantification methods play vital roles in molecular biology.With the development of molecular biology,isothermal amplification of DNA/RNA,as a new molecular biology technology,can be amplified under isothermal condition,it has the advantages of high sensitivity,high specificity,and high efficiency,and has been applied in various fields of biotechnology,including disease diagnosis,pathogen detection,food hygiene and safety detection and so on.This paper introduces the progress of isothermal amplification technology,including rolling circle amplification(RCA),nucleic acid sequence-dependent amplification(NASBA),strand displacement amplification(SDA),loop-mediated isothermal amplification(LAMP),helicase-dependent amplification(HDA),recombinase polymerase amplification(RPA),cross-priming amplification(CPA),and its principle,advantages and disadvantages,and application development are briefly summarized.展开更多
Plant pathogens cause severe losses to crop yields and economic returns in agriculture.Despite plant tissue DNA extraction of typically constituting a preliminary step in nucleic acid-based molecular diagnostics,such ...Plant pathogens cause severe losses to crop yields and economic returns in agriculture.Despite plant tissue DNA extraction of typically constituting a preliminary step in nucleic acid-based molecular diagnostics,such lab-based methods can be time-consuming and arduous to complete many samples.To mitigate these challenges,we developed an inexpensive portable DNA extraction technique that is lightweight and suitable for deployment in sampling locations,such as fields.It includes a DNA extraction device fabricated with a Steel Microneedle Array(SMA)and a simple high-efficiency DNA extraction buffer.As a result,DNA extraction times can be reduced to within~1 min,and the eluted DNA is demonstrated to be suitable for subsequent molecular biological analyses without requiring additional purification.Cross-priming amplification(CPA)technology was first established to detect Phytophthora infestans,which achieves sensitivity attainment of 10^(–7)ng/μL.The detection result can be conveniently estimated with naked-eye visual inspection using fluorescent dsDNA binding dye.CPA was demonstrated to be more feasible than PCR-based approaches and performed well in species-specific and practicability tests.This study elucidates a novel integrated pathogen detection technique coupled with SMA-Device extraction and a modified visual CPA assay to establish and verify various field-based samples infected with multiple pathogens.Altogether,the total sample-toanswer time for pathogen detection was reduced to~1.5 h,making field-based analysis affordable and achievable for farmers or extension workers inside and outside the laboratory.展开更多
T lymphocytes with dedicated suppressor function(Treg)play a crucial role in the homeostatic control of immunity in the periphery.Several Treg phenotypes have now been identified in the CD4 and CD8 T cell populations,...T lymphocytes with dedicated suppressor function(Treg)play a crucial role in the homeostatic control of immunity in the periphery.Several Treg phenotypes have now been identified in the CD4 and CD8 T cell populations, suggesting their down-regulatory function in both human and animal models of autoimmunity,transplantation and tumor immunity.Here we will focus on the CD8 Treg population and their ability to specifically inhibit a pathogenic autoimmune response.This review will detail the current advances in the knowledge of CD8 Treg in the context of antigen specificity,phenotype,MHC restriction,mechanism of action,and priming.Cellular & Molecular Immunology.2005;2(1):11-19.展开更多
基金supported by grants from Jiangsu Higher Education Institution Innovative Research Team for Science and Technology(2021),the Key Technology Program of Suzhou People’s Livelihood Technology Projects(Grant Nos.SKY2021029,SZS2020311)the Open Project of Jiangsu Biobank of Clinical Resources(TC2021B009)the Qing-Lan Project of Jiangsu Province in China(2021,2022).
文摘Nucleic acid(DNA and RNA)detection and quantification methods play vital roles in molecular biology.With the development of molecular biology,isothermal amplification of DNA/RNA,as a new molecular biology technology,can be amplified under isothermal condition,it has the advantages of high sensitivity,high specificity,and high efficiency,and has been applied in various fields of biotechnology,including disease diagnosis,pathogen detection,food hygiene and safety detection and so on.This paper introduces the progress of isothermal amplification technology,including rolling circle amplification(RCA),nucleic acid sequence-dependent amplification(NASBA),strand displacement amplification(SDA),loop-mediated isothermal amplification(LAMP),helicase-dependent amplification(HDA),recombinase polymerase amplification(RPA),cross-priming amplification(CPA),and its principle,advantages and disadvantages,and application development are briefly summarized.
基金supported by the National Key Research and Development Program of the Ministry of Science and Technology(2022YFD1400104)the Talent Cultivation and Development Support Program,China Agricultural University.
文摘Plant pathogens cause severe losses to crop yields and economic returns in agriculture.Despite plant tissue DNA extraction of typically constituting a preliminary step in nucleic acid-based molecular diagnostics,such lab-based methods can be time-consuming and arduous to complete many samples.To mitigate these challenges,we developed an inexpensive portable DNA extraction technique that is lightweight and suitable for deployment in sampling locations,such as fields.It includes a DNA extraction device fabricated with a Steel Microneedle Array(SMA)and a simple high-efficiency DNA extraction buffer.As a result,DNA extraction times can be reduced to within~1 min,and the eluted DNA is demonstrated to be suitable for subsequent molecular biological analyses without requiring additional purification.Cross-priming amplification(CPA)technology was first established to detect Phytophthora infestans,which achieves sensitivity attainment of 10^(–7)ng/μL.The detection result can be conveniently estimated with naked-eye visual inspection using fluorescent dsDNA binding dye.CPA was demonstrated to be more feasible than PCR-based approaches and performed well in species-specific and practicability tests.This study elucidates a novel integrated pathogen detection technique coupled with SMA-Device extraction and a modified visual CPA assay to establish and verify various field-based samples infected with multiple pathogens.Altogether,the total sample-toanswer time for pathogen detection was reduced to~1.5 h,making field-based analysis affordable and achievable for farmers or extension workers inside and outside the laboratory.
文摘T lymphocytes with dedicated suppressor function(Treg)play a crucial role in the homeostatic control of immunity in the periphery.Several Treg phenotypes have now been identified in the CD4 and CD8 T cell populations, suggesting their down-regulatory function in both human and animal models of autoimmunity,transplantation and tumor immunity.Here we will focus on the CD8 Treg population and their ability to specifically inhibit a pathogenic autoimmune response.This review will detail the current advances in the knowledge of CD8 Treg in the context of antigen specificity,phenotype,MHC restriction,mechanism of action,and priming.Cellular & Molecular Immunology.2005;2(1):11-19.
