ExperimentalStudyonDetectionofDeadandLive Mycobacterium Tuberculosis by FlowCytometry
- VernacularTitle:应用流式细胞术鉴定死活结核分枝杆菌的实验研
- Author:
Wei-liang WANG
1
;
Bei XIE
2
;
Fan-rong MENG
2
;
Nan WANG
2
;
Yu YANG
2
;
Zhi-hui LIU
2
;
Shou-yong TAN
2
;
Yan-bin ZHANG
3
Author Information
1. GuangzhouMedicalUniversity，Guangzhou511436，China
2. InstituteofPulmonaryDiseases，GuangzhouChestHospital，Guangzhou 510095，China
3. InternalMedicine-Tuberculosis，GuangzhouChest Hospital，Guangzhou510095，China
- Publication Type:Journal Article
- Keywords:
flowcytometry；Mycobacterium tuberculosis；propidiumiodide；6-carboxyfluoresceindiacetae
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2019;40(3):372-378
- CountryChina
- Language:Chinese
-
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.
