Determination of Baylisascaris schroederi Infection in Wild Giant Pandas by an Accurate and Sensitive PCRCE-SSCP Method 英文参考文献.docVIP

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Determination of Baylisascaris schroederi Infection in Wild Giant Pandas by an Accurate and Sensitive PCRCE-SSCP Method 英文参考文献.doc

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Determination of Baylisascaris schroederi Infection in Wild Giant Pandas by an Accurate and Sensitive PCRCE-SSCP Method 英文参考文献

DeterminationofBaylisascarisschroederiInfectionin WildGiantPandasbyanAccurateandSensitivePCR/CE- SSCPMethod WenpingZhang1,ShangmianYie1,BisongYue2*,JielongZhou3,RenxiongAn1,JiangdongYang4, WangliChen5,ChengdongWang1,LiangZhang1,FujunShen1,GuangyouYang4,RongHou1, ZhiheZhang1* 1TheSichuanKeyLaboratoryofConservationBiologyonEndangeredWildlife,ChengduResearchBaseofGiantPandaBreeding,Chengdu,Sichuan,People’sRepublicof China,2TheSichuanKeyLaboratoryofConservationBiologyonEndangeredWildlife,CollegeofLifeScience,SichuanUniversity,Chengdu,People’sRepublicofChina, 3College of Life Sciences, Southwest Forestry University, Kunming, People’s Republic of China, 4College of Animal Science, Sichuan Agricultural University, Ya’an, People’sRepublicofChina,5TangjiaheNationalNatureReserve,Guangyuan,People’sRepublicofChina Abstract It has been recognized that other than habitat loss, degradation and fragmentation, the infection of the roundworm Baylisascarisschroederi(B.schroederi)isoneofthemajorcausesofdeathinwildgiantpandas.However,theprevalenceand intensityoftheparasiteinfectionhasbeeninconsistentlyreportedthroughamethodthatusessedimentation-floatation followedbyamicroscopeexamination.Thismethodfailstoaccuratelydetermineinfectionbecausetherearemanybamboo residuesand/orfewB.schroederieggsintheexaminedfecalsamples.Inthepresentstudy,weadoptedamethodthatuses PCR and capillary electrophoresis combined with a single-strand conformation polymorphism analysis (PCR/CE-SSCP) to detect B. schroederi infection in wild giant pandas at a nature reserve, and compared it to the traditional microscope approach.ThePCRspecificallyamplifiedasinglebandof279-bpfrombothfecalsamplesandpositivecontrols,whichwas confirmed by sequence analysis to correspond to the mitochondrial COII gene of B. schroederi. Moreover, it was demonstratedthattheamountofgenomicDNAwaslinearlycorrelatedwiththepeakareaoftheCE-SSCPanalysis.Thus, ouradoptedmethodcanreliablydetecttheinfectiousprevalenceandintensityofB.schroederiinwildg