A pyrosequencing-based method for genotyping pathogenic serotypes of S. suis
[摘要] Streptococcussuis(S.suisforshort)cancauseavarietyofinfectionsinpigs,andtheinfectionshavebroughtaboutgreatlossesintheswineindustryandsomecasesofdeathsinhumanbeings.InordertorapidlydiagnoseandcontroltheinfectionsofS.suis,wedesignedapyrosequencing-basedassaytoidentifytheserotypesofS.suis.Intheassay,pyrosequencingisusedtogenotypemostofthepathogenicserotypesofS.suisbydetectingfiveinformativeregionsontheChaperonin60(cpn60)geneandonespecies-specificregiononthe16SrRNAgene,andfurtherafewundistinguishedserotypesbypyrosequencingwerefinelydiscriminatedbymultiplexPCRofserotype-specificfragmentsonthecpsgeneaswellasspecies-specificfragmentsonthe16SrRNAgene.ThroughcarefullydesigningthedispensingorderofdNTPforeachpyrosequencingreaction,theserotypesofS.suiscouldbediscriminatedbyfourpyrosequencingreactionswithinthreehours.Fivereferenceserotypesandthreeclinicalstrainsweresuccessfullydetectedandgenotypedbyourassay.Theresultsindicatedthatourassayisareliable,information-richdiagnosticmethodfortheaccuratedetectionofS.suisserotypes...
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[效力级别] [学科分类] 分析化学
[关键词] [时效性]
