JournalofVisualizedExperimentswww.jove.comCopyright©2011CreativeCommonsAttribution-NonCommercialLicenseJuly2011|53|e3104|Page1of16VideoArticleDNAFingerprintingofMycobacteriumlepraeStrainsUsingVariableNumberTandemRepeat(VNTR)-FragmentLengthAnalysis(FLA)RonaldW.Jensen1,JasonRivest1,WeiLi1,VaralakshmiVissa11DepartmentofMicrobiology,ImmunologyandPathology,ColoradoStateUniversityCorrespondenceto:VaralakshmiVissaatvvissa@colostate.eduURL:http://www.jove.com/video/3104DOI:doi:10.3791/3104Keywords:Immunology,Issue53,Mycobacteriumleprae,leprosy,biopsy,STR,VNTR,PCR,fragmentlengthanalysisDatePublished:7/15/2011Citation:Jensen,R.W.,Rivest,J.,Li,W.,Vissa,V.DNAFingerprintingofMycobacteriumlepraeStrainsUsingVariableNumberTandemRepeat(VNTR)-FragmentLengthAnalysis(FLA).J.Vis.Exp.(53),e3104,doi:10.3791/3104(2011).AbstractThestudyofthetransmissionofleprosyisparticularlydifficultsincethecausativeagent,Mycobacteriumleprae,cannotbeculturedinthelaboratory.Theonlysourcesofthebacteriaareleprosypatients,andexperimentallyinfectedarmadillosandnudemice.Thus,manyofthemethodsusedinmodernepidemiologyarenotavailableforthestudyofleprosy.DespiteanextensiveglobaldrugtreatmentprogramforleprosyimplementedbytheWHO1,leprosyremainsendemicinmanycountrieswithapproximately250,000newcaseseachyear.2TheentireM.lepraegenomehasbeenmapped3,4andmanylocihavebeenidentifiedthathaverepeatedsegmentsof2ormorebasepairs(calledmicro-andminisatellites).5ClinicalstrainsofM.lepraemayvaryinthenumberoftandemrepeatedsegments(shorttandemrepeats,STR)atmanyoftheseloci.5,6,7Variablenumbertandemrepeat(VNTR)5analysishasbeenusedtodistinguishdifferentstrainsoftheleprosybacilli.Someofthelociappeartobemorestablethanothers,showinglessvariationinrepeatnumbers,whileothersseemtochangemorerapidly,sometimesinthesamepatient.WhilethevariabilityofcertainVNTRshasbroughtupquestionsregardingtheirsuitabilityforstraintyping7,8,9,theemergingdatasuggestthatanalyzingmultipleloci,whicharediverseintheirstability,canbeusedasavaluableepidemiologicaltool.Mul...
