Porcine proliferative enteropathy(PPE),an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis,is commonly associated with diarrhea and reduced weight g...Porcine proliferative enteropathy(PPE),an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis,is commonly associated with diarrhea and reduced weight gain in growing pigs widespread.An accurate method for detecting L.intracellularis is particularly important for preventing and controlling PPE.Heat shock protein 60(Hsp60)is an immunodominant bacterial antigen found in all eukaryotic and prokaryotic organisms.Thus,the purpose of the current investigation was to produce a novel L.intracellularis Hsp60 monoclonal antibody(mAb)useful for immunodiagnostics.Three hybridomas secreted anti-Hsp60 termed 3E5,4E2,and 9G6 were generated,and the titers of ascitic fluids of 3E5,4E2,9G6 were 1:1024000,1:2048000 and 1:2048000,respectively.The Western blotting analysis demonstrated that recombinant Hsp60(rHsp60)was recognized by mAbs 3E5,4E2 and 9G6.Subsequently,analyses of specificity showed all the mAbs were highly specific to L.intracellularis while could not significantly react with other enteric bacteria commonly found in the ileum of pigs,such as Escherichia coli,Salmonella Choleraesuis,Salmonella Typhimurium,and Brachyspira hyodysenteriae.Furthermore,the mAbs were useful for detecting L.intracellularis in the infected monolayer cells and histological sections of the ileum from PPE-affected pigs.Our research will provide a foundation for the development of immunological diagnostic tests.展开更多
The significance of early detection and isolation of infected individuals,along with the quantitative assessment of antibodies against the virus,has gained widespread recognition during the ongoing covid-19 pandemic.T...The significance of early detection and isolation of infected individuals,along with the quantitative assessment of antibodies against the virus,has gained widespread recognition during the ongoing covid-19 pandemic.This necessitates the development of cost-effective,user-friendly,decentralized testing methods characterized by both high sensitivity and specificity.In this article,we present a comprehensive review of an innovative,low-cost rapid decentralized immunoassay technology,applicable across various diagnostic and quantitative testing scenarios.Distinguishing itself from conventional immunoassay technologies,this method is featured with mechanically enhanced specificity without compromising sensitivity.We delve into the basic principle of the technology and a comparative analysis of this technology in relation to other immunodiagnostic methods,highlighting its potential applications in a wide spectrum of diagnostic tests.展开更多
基金supported by the National Key Research and Development Program of China(2021YFD1800400)the National Natural Science Foundation of China(31872480)+1 种基金the Jiangsu Agriculture Science and Technology Innovation Fund,China(CX(19)2020)the Priority Academic Program Development of Jiangsu Higher Education Institutions,China(PAPD).
文摘Porcine proliferative enteropathy(PPE),an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis,is commonly associated with diarrhea and reduced weight gain in growing pigs widespread.An accurate method for detecting L.intracellularis is particularly important for preventing and controlling PPE.Heat shock protein 60(Hsp60)is an immunodominant bacterial antigen found in all eukaryotic and prokaryotic organisms.Thus,the purpose of the current investigation was to produce a novel L.intracellularis Hsp60 monoclonal antibody(mAb)useful for immunodiagnostics.Three hybridomas secreted anti-Hsp60 termed 3E5,4E2,and 9G6 were generated,and the titers of ascitic fluids of 3E5,4E2,9G6 were 1:1024000,1:2048000 and 1:2048000,respectively.The Western blotting analysis demonstrated that recombinant Hsp60(rHsp60)was recognized by mAbs 3E5,4E2 and 9G6.Subsequently,analyses of specificity showed all the mAbs were highly specific to L.intracellularis while could not significantly react with other enteric bacteria commonly found in the ileum of pigs,such as Escherichia coli,Salmonella Choleraesuis,Salmonella Typhimurium,and Brachyspira hyodysenteriae.Furthermore,the mAbs were useful for detecting L.intracellularis in the infected monolayer cells and histological sections of the ileum from PPE-affected pigs.Our research will provide a foundation for the development of immunological diagnostic tests.
基金funded by the Singapore Ministry of Education Academic Research Funds Tier 2(MOE-T2EP50220-0015)the Ministry of Education under the Research Centres of Excellence programme,the Ministry of Education,Decentralized GAP Funding(TAP2002020-05-18)+1 种基金NUS Efforts Against COVID-19(NUSEAC)FundingNational Research Foundation(NRF)of Singapore,through its NRF Investigator-ship Programme(NRF-NRFI2016-03).
文摘The significance of early detection and isolation of infected individuals,along with the quantitative assessment of antibodies against the virus,has gained widespread recognition during the ongoing covid-19 pandemic.This necessitates the development of cost-effective,user-friendly,decentralized testing methods characterized by both high sensitivity and specificity.In this article,we present a comprehensive review of an innovative,low-cost rapid decentralized immunoassay technology,applicable across various diagnostic and quantitative testing scenarios.Distinguishing itself from conventional immunoassay technologies,this method is featured with mechanically enhanced specificity without compromising sensitivity.We delve into the basic principle of the technology and a comparative analysis of this technology in relation to other immunodiagnostic methods,highlighting its potential applications in a wide spectrum of diagnostic tests.
