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:1005-2224(2007)03-0192-0416SrRNAPCR1,2a,2a,2b:1.,325027;2.ab,310003:E2mail:shangsq33@mail.hz.zj.cn()2003208200521216SrRNA49(CSF)DNA;DNA,CSF(1)PCR(FQ2PCR)4917,3417%(17/49),1012%(5/49),(P0101)(2)17FQ2PCRDNA,DNA,Ct(),Ct,DNA,(3)FQ2PCRCSF5(4)2FQ2PCR,Ct1719,CSF;Ct3118,PCR,,,;;:R72:AArapiddiagnosisofpurulentmeningitisinchildrenby16SrRNAFQ2PCRinCSF.YANGZu2qin,SHANGShi2qiang,WUYi2dong,etal.YuYingChildren’sHospitalAffiliatedtoWenzhouMedicalCollege,Wenzhou325027,ChinaAbstractObjectiveToexploreanewtechniqueofrapiddiagnosisofpurulentmeningitisinchildren.MethodsCe2rebrospinalfluid(CSF)specimensfrom49casesofsuspectedpurulentmeningitisweredetectedby16SrRNAFQ2PCRintheChildrenHospitalAffiliatedtoZhejiangUniversityfromAugust,2003toDecember,2005.Thecopynumbersofbac2terialDNAinCSFweredetected,andthebacterialculturesofCSFweredoneascontrolsatthesametime.Results(1)Ofthe49cases,17werepositivebyFQ2PCR,thepositiverate(3417%)(17/49)wassignificantlyhigherthanthatofCSFculture1012%(5/49)(P0101).(2)ThecopynumbersofbacterialDNAinCSFweredeterminedamongtheseventeenFQ2PCRpositivespecimens,andtheywerefoundtohavepositivecorrelationwiththestateofillnessinchil2dren;thenumberofCtfoundnegativecorrelationwiththeillness.ThelowertheCtnumbers,thehighertheDNAcopynumbers,andtheprognosisofchildrenwouldbeworse.(3)Ofthe49samples,only5caseswerebothpositivebyFQ2PCRandroutingbacterialcultureofCSF.(4)TwoproductsofFQ2PCRinCSFweresequenced.BacteriumcoliwereshowninthesequenceresultofCt1719;itcorrespondedwiththeresultofCSFculture.SequencingfailedinCt3118.ConclusionFQ2PCRwithhigherspecificityandsensitivity,demandinglessCSF,israpidandreliablefordiagnosingpurulentmen2ingitisinchildren.Soithasconsiderablevalueofapplication.KeywordsFluorescencequantitativepolymerasechainreaction;PurulentMeningitis;CSF(),,4916SrRNAPCR,DNA,CSF,,11.11012,6,6172912007322323,12,11(12)1.220032082005212(ICU)15([1])(130d24,1310,415,233,485,9101,11131)49,2mL,1mL,1mLDNA22.1PCR2.1.1PE25700PCR,SIGMA1215K;dNTP,TaqTakara2.1.2TaqmanGenbank16SrRNA,MegAlign,,,:TGTGTAGCGGTGAAAT2GCG6902708;CATCGTTTACGGCGTGGAC8292811;(UnProbe)TCTAATCCTGTTTGATCCCCACG80027782.1.3PCR,Ep2pendorfDNA,50L(10mmol/LTri2HClpH716,5mmol/LEDTA,015%SDS),10min,,4L,ABI5700,PCR2.1.4PCR944min,9420s6060s,40PCRATCC11775ATCC25923;DNA4,/L1031041051062.21mL,20mL,,3515,3d,24h,VITEK26022.3SPSS11152,P01012.4CSFDNAPCR2CSFDNAPCR40L,33.116SrRNAFQ2PCRFQ2PCRUnProbe(1,2),Ct()1412425108EBDNA16SrRNAFQ2PCR,3.216SrRNAPCR100107/L,RnvsCyclesCt,10,2,Ct3812;,Ct403.3FQ2PCRbaseline0102,Ct35FQ2PCR,FQ2PCR17,3417%(17/49),5,1012%(5/49),2FQ2PCRCSF,2=101481,P=01005,P0101,5CSF,FQ2PCR;12FQ2PCR,CSF,,;32FQ2PCR,CSF3.4FQ2PCRCtDNACt,Ct,DNA,(3)1UnProbeUnProbeUnProbeCt1)+17.281)+20.431)+19.621)+22.151)+15.981)+18.26+15.68+19.68+20.28+23.78+23.09+20.42+23.06+17.59+14.58+16.21+19.26+23.47:1)391200732232UnProbeUnProbeUnProbeCt1)+14.241)+17.851)+14.951)+22.751)+17.261)+21.58+16.58+18.7+22.06+25.08+22.09+17.26+17.24+19.58+18.57+21.08+23.06:1);Ct40,Ct40,baseline01023FQ2PCRCSFDNACtFQ2PCRDNA(copy/L)Ct1+7.1210618.52+8.3210425.83+3.1610328.94+3.7810426.55+2.5610717.96+-7.1110232.67+-5.5610233.18+-9.3710134.39+-4.2210233.810+-4.8110232.211+-6.6310233.512+-1.3110231.813+-5.0110230.414+-7.1110232.615+-1.8910234.216+-2.7610231.917+-8.6510232.33.5CSF1Ct,1719,CSF,,FQ2PCRCt,CSF;1,Ct3118,DNA,,CtCSF4Shang[2]200116SrRNAPCR;200316S223SrRNADNA[3],200416SrRNAPCR[426],Feuillet[7]16SrRNA16SrRNARNADNA,,,PCR,[8]:Taq523,53,(R)(Q),RQ,,,RFQ2PCR,23h,,49:FQ2PCR3417%(17/49),CSF1012%(5/49),,P0101FQ2PCR17FQ2PCRCSF5,12,12:(1)CSFDNA;(2)(3),;FQ2PCRFQ2PCR,FQ2PCRCSFFQ2PCRCSFDNA,Ct,,Ct,DNA,,CSFDNA16SrRNAPCRCSFDNA,;PCR,,[1],[M].//..5.:49120073223,2002:3952396.[2]ShangS,ChenZ,YuX.DetectionofbacterialDNAbyPCRandreversehybridizationinthe16SrRNAgenewithparticularrefer2encetoneonatalsepticemia[J].ActaPaediatr,2001,90(2):1792183.[3]ShangSQ,FUJF,DongG,etal.EstablishmentandanalysisofspecificDNApatternsin16S223SrRNAgenespacerregionsfordifferentbacteria[J].ChinMedJ,2003,116(1):1292133.[4]ShangS,ChenG,WuY,etal.RapiddiagnosisofbacterialsepsiswithPCRamplificationandmicroarrayhybridizationin16SrRNAgene[J].PediatrRes.2005,58(1):1432148.[5],,,.16SrRNAPCR[J].,2004,42(9):6632667.[6],,,.16SrRNA[J].,2005,23(3):1872190.[7]FeuilletL,CarvajalJ,SudreI,etal.Firstisolationofbacteroidesthetaiotaomicronfromapatientwithacholesteatomaandexperi2encingmeningitis[J].JClinMicrobiol,2005,43(3):146721469.[8]PasSD,FriesE,DeManRA,etal.Developmentofaquantitativereal2timedetectionassayforhepatitisBvirusDNAandcompari2sonwithtwocommercialassays[J].JClinMicrobiol,2000,38(8):289722901.(2006-09-022006-12-04)(:)1:,810001E2mail:xnmayanyan@126.com,18,10,3200220821210,,,10min8,,,,1h,,,,,12h,37d,3,,,,,5min,25d1,CT,D3,,,,,:T37,,,:3,,(MRA)(DSA),(AHC),2.5mg,2,,:(AHC),AHCAicardi[1]1987:(1)18;(2),;(3),;(4),,Todd,,MRIMRA,,,,:(1);(2);(3)22(MELAS);(4),,AHC,,[1]AminianA,StrashunA,RoseA.Alternatinghemiplegiaofchildhood:studiesofregionalcerebralbloodflowusing99mTchexamethylpropyleneamineoximesingle2photonemissioncomputedtomography[J].AnnNeurol,1993.33:43247.(2006-11-102007-