北方某地区牛布鲁菌病流行病学调查及布鲁菌多重pcr检测方法建立

《北方某地区牛布鲁菌病流行病学调查及布鲁菌多重pcr检测方法建立》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、摘要为了解北方某地区牛群中布鲁菌病的疫情，采取更为有效的防控措施，本文利用虎红平板凝集试验（RBT）、试管凝集试验（SAT）和间接酶联免疫吸附试验（I-ELISA）随机对该地区牧场及散养牛群进行了布鲁菌病流行病学调查，同时对以上三种方法的检测水平进行了比较；根据GenBank已发表的布鲁菌基因序列计合成三对特异性引物，建立了同时检测布鲁菌属及牛、羊种布鲁菌的诊断鉴别性多重PCR方法，并对其进行了初步应用。用上述三种方法随机对北方某地区牧场和散养牛的950份血清进行布鲁菌病血清抗体检测，表明该地区布鲁菌病流行

2、情况较为严重，但不同群体阳性率差异较大，牧场阳性率最高达50%，最低阳性率仅为4.6%，种牛场未检出阳性血清，散养奶牛阳性率为39.3%；RBT检出阳性血清105份，阳性率为15.8%，SAT检出阳性血清128份，阳性率为19.2%，I-ELISA检出阳性血清262份，阳性率为27.6%，结果表明I-ELISA阳性率最高，SAT次之，RBT最低。所建立多重PCR方法的敏感性可达100pg左右，且具有很好的特异性与重复性，对7株参考株葡萄球菌26001等的PCR扩增均为阴性，初步临床样品应用表明该方法可用于乳样

3、、阴道及粪便棉拭的检测。关键词：布鲁菌病；流行病学调查；多重PCR；应用BovineBrucellosisSerologicalInvestigationinaRegionofNorthChinaandFormationofMulti-PCRDetectingforBrucellosisAbstactTounderstandthesituationofbrucellosisinaregionofnorthChinaandapplythemoreeffectiveprecautions,thisstudyapp

4、liedepidemiologicalsurveystochasticallyinthegrazingfarmsanddistributivecattlegroupsinthisregion,usingRoseBengalPrecipitationTest(RBT),SerumAgglutinationTest(SAT)andIndirectEnzymeLinkedImmunosorbentAssay(I-ELISA),andcomparedthethreeexperiments,respectively.B

5、asedonthebrucellosisgenesequencepublishedonGenBank,specializedprimerwassynthesized,multi-PCRidentificationanddiagnosisofBrucellaandbrucellosisoncowsandsheepwereestablishedandprimaryapplicationtestwasalreadyconducted.Stochasticallyusingthethreemethodsabove,9

6、50serumsamplesfromthegrazingfarmsanddistributivecattlegroupsinaregionofnorthChinawereinspected.Resultsshowedthattheepidemicsituationofbrucellosisisrelativelyserious.However,positiveratesgreatlyvaryamonggroupswiththehighest50%andthelowest4.6%inthegrazingfarm

7、sandnobrucellosiswerefoundinthebreedingbullfarmsandthepositiverateindistributivecattlegroupswas39.3%;RBTtested105serumsamplesandthepositiverateis15.8%;SATtested128serumsamplesandthepositiveratewas19.2%;I-ELISAtested262serumsamplesandthepositiveratewas27.6%.

8、ResultsshowtheI-ELISAenjoysthegreatestpositiverate,higherthanSATandRBT.RBTranksasthelast.Thesensitivityofmulti-PCRcouldreachapproximately100pg,possessingexcellentspecificityandrepeatability.PCRamplific