简介:Purifiedproteinderivative(PPD)skintestsoftenyieldpoorspecificity,sothattodevelopnewserologicalantigensfordistinguishingbetweenMycobacteriumtuberculosisinfectionandBacilleCalmette-Guerin(BCG)vaccinationisapriority,especiallyfordevelopingcountrieslikeChina.Wepredictedtheantigenicityforselectedopenreadingframes(ORFs)basedonthegenomesequencesofM.tuberculosisH37RvandM.bovisBCG,aswellastheirfunctionsanddifferencesofexpressionunderdifferentstimulus.ThecandidateORFswereclonedfromH37RvsequencesandexpressedasrecombinantproteinsinEscherichiacoli.Westudiedtheserodiagnosticpotentialof11purifiedrecombinantsbyusingenzyme-linkedimmunosorbentassay(ELISA)andinvolvingacohortcomposedof58TBpatients(34malesand24females),8healthyvolunteersand50PPD-negativeindividualsbeforeandafterBCGvaccination.Forallthe11antigens,themedianODvaluesfortheserafromTBpatientswerestatisticallysignificantlyhigherthanthoseforPPD-negativeindividualsbeforeorafterBCGvaccination(P<0.01).Theyhadatleast92%specificityinhealthycontrolsandsixseroantigens(Rv0251c,Rv1973,Rv2376c,Rv2537c,Rv2785candRv3873A)wereneverreportedwithseroantigenicitiespreviously.Thustheapproachcombiningcomparativegenomics,bioinformaticsandELISAtechniquescanbeemployedtoidentifynewseroantigensdistinguishingM.tuberculosisinfectionfromBCGvaccination.
简介:一个新奇高产量的系统,为分离和反应(4SR)叫了叠的片胶化系统,为DNA/RNA和蛋白质/肽的分析被开发。系统提供利用叠的片胶化的性质的一条新奇三维的胶化电气泳动途径。它同时允许多重样品反应以及被分开,出现一二维(mxn)样品装载系统。为这个目的,包含井(在这篇论文的100口井)的可变数字的高产量的多微的容器(MMV)被使用了,它用25公里做的是方形尺寸的polyacrylamide胶化。在electrophoretic分离以后,而且,包含一件需要的样品的片胶化能容易被移开并且继续到下一步。不同生物反应以及产品的连续分离有效地被执行处理DNA/RNA和蛋白质/肽。它证明这个系统有多种潜力被发展。