ExperimentalStudyonDetectionofDeadandLive Mycobacterium Tuberculosis by FlowCytometry / 中山大学学报(医学科学版)
Journal of Sun Yat-sen University(Medical Sciences)
; (6): 372-378, 2019.
Article
in Zh
| WPRIM
| ID: wpr-817782
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WPRO
ABSTRACT
@#:【Objective】To provide basic experimental basis for quantitative detection and analysis of antibiotic susceptibilityandheteroresistancein Mycobacterium tuberculosis byflowcytometry,weinvestigatedtheperformanceof propidiumiodide(PI)and6-carboxyfluoresceindiacetae(6-cFDA)inidentifyingdeadorlivingMycobacterium tubercu⁃ losis.【Methods】PIand6-cFDAwereusedtostain10strainsofpureculturedliving Mycobacterium tuberculosis suspen⁃ sion with turbidity of 1 McClell and the corresponding suspension with turbidity of 1 McClell heated at 85℃ for 30 minutes,respectively.Themeanfluorescenceintensity(MFI)wasmeasuredbyflowcytometry.TheMFIboundaryvalue fordistinguishingdeadandlivingMycobacterium tuberculosisweredeterminedaccordingtoreceiveroperatingcharacteristic (ROC)curves.Thesameexperimentswerecarriedoutwithother10strainsofliving Mycobacterium tuberculosis suspen⁃sionand10dead Mycobacterium tuberculosis suspension,andtheMFIthresholdwasusedtodeterminetheviabilityof bacteria.【Results】(1)IntheexperimentofdeterminingMFIboundaryvalue,theMFIof10strainsofdeadbacteria suspensionstainedwithPIwasnon-normaldistribution,themedianvaluewas2459,andtheMFIof10strainsofliving bacteriasuspensionstainedwithPIwasnormaldistribution,themeanvaluewas426±180.TheMFIboundaryvalueof deadbacteriastainedwithPIwas1329.TheMFIof10strainsofdeadbacteriaand10strainsoflivingbacteriasuspen⁃ sionstainedwith6-cFDAwasnormaldistribution,andthemeanvaluewas49±4and7144±4511,respectively.The MFIthresholdof6-cFDAstainingforlivingbacteriawas1021.(2)TheMFIthresholdsforidentifyingdeadbacteriaby PIstainingandtheMFIthresholdsforidentifyinglivingbacteriaby6-cFDAstainingwereusedtodetect10otherliving bacterialsuspensionsand10deadbacterialsuspensions.Theaccuracy,sensitivity,specificity,Yondeindex,positive predictivevalueandnegativepredictivevaluewere95%and100%,1.00and1.00,0.90and1.00,0.90and1.00,0.91 and1.00,1.00and1.00,respectively.【Conclusions】PIstainingcandetectdeadMycobacterium tuberculosisand6-cFDA stainingcandetectliving Mycobacterium tuberculosis.Identificationof Mycobacterium tuberculosis activitybasedonPIand 6-cFDAstainingwillhavebroadapplicationprospects.
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WPRIM
Language:
Zh
Journal:
Journal of Sun Yat-sen University(Medical Sciences)
Year:
2019
Type:
Article