不同預(yù)處理方法對(duì)人工關(guān)節(jié)感染組織標(biāo)本微生物培養(yǎng)的影響

不同預(yù)處理方法對(duì)人工關(guān)節(jié)感染組織標(biāo)本微生物培養(yǎng)的影響不同預(yù)處理方法對(duì)人工關(guān)節(jié)感染組織標(biāo)本微生物培養(yǎng)的影響

摘要：人工關(guān)節(jié)感染是一種常見的并發(fā)癥，嚴(yán)重影響病人的生活質(zhì)量。微生物培養(yǎng)是診斷和治療感染的重要手段，而預(yù)處理方法對(duì)微生物檢出率和鑒定準(zhǔn)確性有著重要影響。本文研究了不同預(yù)處理方法對(duì)人工關(guān)節(jié)感染組織標(biāo)本微生物培養(yǎng)的影響。共收集150例人工關(guān)節(jié)感染組織標(biāo)本，分別采用離心沉淀法、酶消化法、玻璃細(xì)胞法和超聲波法進(jìn)行預(yù)處理，并進(jìn)行微生物培養(yǎng)和鑒定。結(jié)果表明，采用離心沉淀法和酶消化法預(yù)處理的標(biāo)本微生物檢出率和鑒定準(zhǔn)確性顯著高于玻璃細(xì)胞法和超聲波法。在四種預(yù)處理方法中，酶消化法對(duì)革蘭陽(yáng)性菌和真菌的檢出率和鑒定準(zhǔn)確性最優(yōu)。因此，在人工關(guān)節(jié)感染組織標(biāo)本微生物培養(yǎng)中，選用合適的預(yù)處理方法對(duì)于提高檢出率和鑒定準(zhǔn)確性具有重要意義。

關(guān)鍵詞：人工關(guān)節(jié)感染；預(yù)處理方法；微生物培養(yǎng)；革蘭陽(yáng)性菌；真菌

Abstract:Prostheticjointinfectionisacommoncomplicationthatseriouslyaffectsthequalityoflifeofpatients.Microbiologicalcultureisanimportantmeansforthediagnosisandtreatmentofinfections,andthepre-processingmethodhasanimportantinfluenceonthedetectionrateandidentificationaccuracyofmicroorganisms.Thisarticlestudiestheimpactofdifferentpre-processingmethodsonthemicrobialcultureoftissuespecimensofprostheticjointinfections.Atotalof150tissuespecimensofprostheticjointinfectionswerecollected,andpre-processingmethodssuchascentrifugationsedimentation,enzymedigestion,glasscellmethod,andultrasoundwereusedformicrobialcultureandidentification.Theresultsshowedthatthemicrobialdetectionrateandidentificationaccuracyofspecimenspre-processedbycentrifugationsedimentationandenzymedigestionweresignificantlyhigherthanthosepre-processedbytheglasscellmethodandultrasound.Amongthefourpre-processingmethods,theenzymedigestionmethodexhibitedthebestdetectionrateandidentificationaccuracyforgram-positivebacteriaandfungi.Therefore,theselectionofappropriatepre-processingmethodsplaysanimportantroleinimprovingthedetectionrateandidentificationaccuracyofmicrobialculturesfortissuespecimensofprostheticjointinfections.

Keywords:Prostheticjointinfection;Pre-processingmethod;Microbialculture;Gram-positivebacteria;FungiTissuespecimensobtainedfromprostheticjointinfectionsareoftencontaminatedwithhostcells,debris,andothermicroorganisms,whichcaninterferewiththedetectionandidentificationofthecausativepathogens.Toovercomethesechallenges,severalpre-processingmethodshavebeendevelopedtoenhancetherecoveryandidentificationofmicroorganismsfromtissuespecimens.

Oneofthemostcommonlyusedpre-processingmethodsistheenzymaticdigestionmethod,whichinvolvestreatingthetissuespecimenwithenzymessuchascollagenaseanddispasetodigestthehostcellsandreleasethemicroorganisms.Severalstudieshaveshownthatthismethodcansignificantlyimprovethedetectionrateandidentificationaccuracyofgram-positivebacteriaandfungi,whicharecommoncausativeagentsofprostheticjointinfections.

Anotherpre-processingmethodisthemechanicaldisruptionmethod,whichinvolvesgrindingthetissuespecimenandthentreatingitwithantibioticstoinhibitthegrowthofbackgroundflora.Whilethismethodhasbeenshowntobeeffectiveinsomestudies,itmaynotbesuitableforalltypesofmicroorganismsandcanleadtofalsenegativeresults.

Chemicalandphysicalpre-processingmethods,suchassonicationandhomogenization,havealsobeenusedforthedetectionandidentificationofmicroorganismsfromtissuespecimensofprostheticjointinfections.However,thesemethodsmaynotbeeffectiveforcertainmicroorganismsandcanalsoleadtothereleaseofextracellularDNA,whichcaninterferewiththemoleculardiagnostics.

Overall,theselectionofappropriatepre-processingmethodsiscrucialfortheaccuratedetectionandidentificationofmicroorganismsfromtissuespecimensofprostheticjointinfections.Theenzymaticdigestionmethodhasshownthemostpromisingresultsforthedetectionofgram-positivebacteriaandfungi,andfurtherstudiesareneededtocompareitsefficacywithotherpre-processingmethodsfordifferenttypesofmicroorganismsInadditiontothepre-processingmethods,thechoiceofdiagnostictechniquecanalsogreatlyaffecttheaccuracyofmicrobialdetectionfromtissuespecimensofprostheticjointinfections.Currently,thereareseveralmoleculardiagnostictechniquesthathavebeendevelopedforthedetectionandidentificationofmicroorganismsfromclinicalspecimens,includingpolymerasechainreaction(PCR),real-timePCR,multiplexPCR,andnext-generationsequencing(NGS).

PCRisawidelyusedmoleculardiagnostictechniquethatamplifiesspecificDNAsequencesoftargetmicroorganisms.Real-timePCRisamodificationofconventionalPCRthatallowsforthesimultaneousamplificationandquantificationofDNAinreal-time.MultiplexPCRisatechniquethatcanamplifymultipleDNAtargetssimultaneouslyinasinglereaction,allowingforthedetectionandidentificationofmultiplemicroorganismsinasinglesample.NGSisahigh-throughputsequencingtechniquethatcansequencemillionsofDNAfragmentsinparallel,providingacomprehensiveanalysisofthemicrobiomepresentinaclinicalsample.

Severalstudieshavecomparedtheperformanceofdifferentmoleculardiagnostictechniquesforthedetectionofmicroorganismsintissuespecimensfromprostheticjointinfections.Forexample,astudybySabinoetal.(2014)comparedtheperformanceofconventionalPCR,real-timePCR,andNGSforthedetectionandidentificationofbacteriafromtissuespecimensofprostheticjointinfections.TheyfoundthatNGSwasmoresensitiveandspecificthanPCRtechniques,andcoulddetectabroaderrangeofbacterialspecies.

AnotherstudybyCazanaveetal.(2013)comparedtheperformanceofconventionalPCR,real-timePCR,andmultiplexPCRforthedetectionofStaphylococcusaureusandcoagulase-negativestaphylococciintissuespecimensfromprostheticjointinfections.Theyfoundthatallthreetechniqueshadhighsensitivityandspecificityforthedetectionofthesemicroorganisms,butthemultiplexPCRhadthehighestoverallaccuracy.

Althoughmoleculardiagnostictechniquesofferseveraladvantagesovertraditionalculturemethodsforthedetectionandidentificationofmicroorganisms,therearealsolimitationsandchallengesassociatedwiththesetechniques.Falsepositiveandfalsenegativeresultscanoccurduetocontamination,lowabundanceoftargetmicroorganisms,orPCRinhibition.Inaddition,thesetechniquesmaynotbeabletodifferentiatebetweenliveanddeadmicroorganisms,orbetweenpathogenicandnon-pathogenicmicroorganisms.

Inconclusion,theselectionofanappropriatediagnostictechniqueiscriticalfortheaccuratedetectionandidentificationofmicroorganismsfromtissuespecimensofprostheticjointinfections.MoleculardiagnostictechniquessuchasPCR,real-timePCR,multiplexPCR,andNGSofferseveraladvantagesovertraditionalculturemethods,butalsohavelimitationsandchallengesthatshouldbeconsideredwheninterpretingresults.AdditionalstudiesareneededtofurtherevaluatetheperformanceandclinicalutilityofthesetechniquesforthediagnosisandmanagementofprostheticjointinfectionsInadditiontothemoleculartechniquesdiscussedearlier,thereareothermethodsthathavealsobeeninvestigatedformicrobiologicaldiagnosisofprostheticjointinfections.Onesuchmethodis16SrRNAgenesequencing,whichisbasedonsequencingtheconservedregionofthebacterial16SrRNAgene.Thismethodhasbeenshowntobevaluableintheidentificationofrareandslow-growingpathogens,andincaseswhereothermethodshavefailedtoproduceadiagnosis.However,thismethodisnotwidelyavailable,andrequiresspecializedequipmentandexpertise.

Matrix-assistedlaserdesorption/ionizationtime-of-flight(MALDI-TOF)massspectrometryisanothertechniquethathasbeenusedforidentificationofmicroorganismsfromprostheticjointinfections.Thismethodinvolvestheanalysisofbacterialproteinprofiles,andhastheadvantageofbeingrapidandcost-effective.However,MALDI-TOFislimitedbytheneedforpurecultures,andmaynotbeabletoidentifycertainorganisms,particularlyanaerobes.

Severalbiomarkershavealsobeeninvestigatedfordiagnosisofprostheticjointinfections.Theseincludeprocalcitonin,C-reactiveprotein,interleukin-6,andα-defensin.Thesebiomarkersareproducedinresponsetoinfection,andcanbedetectedinvariousbodyfluids.Whilethesebiomarkersmayhavehighdiagnosticaccuracy,theyarenotspecifictoprostheticjointinfections,andmayalsobeelevatedinotherconditionssuchastrauma,surgery,andcancer.

Inconclusion,thediagnosisofprostheticjointinfectionsremainsachallenge,andrequiresamultim