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改良型环介导等温扩增法检测金黄色葡萄球菌的研究.docx

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改良型环介导等温扩增法检测金黄色葡萄球菌的研究 Abstract： Staphylococcusaureusisacommonpathogenicbacteriumthatcancauseavarietyofinfectiousdiseases.TherapiddetectionofStaphylococcusaureusisofgreatsignificanceforthepreventionandcontrolofclinicalinfections.Theimprovedloop-mediatedisothermalamplificationmethodbasedonthecircularizedmolecularbeacon(cMB-LAMP)wasusedtodetectStaphylococcusaureusinthisstudy.TheresultsshowedthatthecMB-LAMPmethodhadgoodspecificity,sensitivityandrepeatability,whichissuitablefortherapiddetectionofStaphylococcusaureus. Introduction： StaphylococcusaureusisaGram-positivebacteriumthatiscommonlyfoundontheskinandmucousmembranes.Itisamajorpathogenthatcancauseskininfections,pneumonia,sepsis,foodpoisoningandotherinfectiousdiseases.Inrecentyears,theincidenceofStaphylococcusaureusinfectionshasbeenincreasingyearbyyear,andthemortalityrateofpatientswithsevereinfectionsisalsoincreasingduetotheemergenceofdrug-resistantstrains.Therefore,itisofgreatsignificancetodeveloparapidandaccuratemethodforthedetectionofStaphylococcusaureus. Theloop-mediatedisothermalamplification(LAMP)methodisarapidandefficientnucleicacidamplificationmethodthatcanamplifytargetDNAataconstanttemperature.However,theconventionalLAMPmethodhassomelimitations,suchashighcost,lowspecificity,anddifficultyinvisualdetectionofamplificationproducts.Inrecentyears,manymodifiedLAMPmethodshavebeendevelopedtoovercometheseshortcomings. Circularizedmolecularbeacon(cMB)isakindofmolecularbeaconthathasbeencircularizedbynucleicacidligation.Unlikelinearmolecularbeacons,cMBscanformaclosed-loopstructure,whichcaneffectivelyreducetheinterferenceofnon-specificDNAamplification.Inaddition,cMBscanbindtotargetDNAwithhighspecificityandsensitivity.Therefore,cMBshavebeenwidelyusedinvariousnucleicaciddetectionmethods. Inthisstudy,animprovedLAMPmethodbasedoncMB(cMB-LAMP)wasdevelopedtodetectStaphylococcusaureus.Thespecificity,sensitivityandrepeatabilityofthecMB-LAMPmethodwereevaluated. MaterialsandMethods： 1.DesignofprimersandcMBs Basedontheconservedseque