?url_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Adc&rft.relation=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F10593%2F&rft.title=Optimization+of+rTDMH+as+a+Reagent+Toward+Improving+the+Sensitivity+of+the+RT-PCR+Based+Diagnosis+for+Mycobacterium+Tuberculosis&rft.creator=Bhatti%2C+Alexandra&rft.description=Current+diagnostic+tools+being+used+for+tuberculosis+lack+the+speed+and+sensitivity+necessary+to+successfully+combat+the+current+tuberculosis+epidemic.++Real-time+Polymerase+Chain+Reaction%2C+RT-PCR%2C+can+provide+the+rapid+and+specific+diagnosis+that+is+currently+in+demand+in+the+global+community.++Its+disadvantage+is+that+due+to+the+waxy+and+robust+nature+of+the+M.+tuberculosis+membrane%2C+not+enough+genomic+DNA+is+present+to+provide+for+amplification+in+a+RT-PCR.+It+was+previously+found+in+our+laboratory+that+hydrolysis+of+one+of+abundant+glycolipid+of+mycobacterial+envelope%2C+Trehalose%2C+6%2C6%E2%80%99-dimycolate%2C+by+a+recombinant+TDM-specific+hydrolase+caused+rapid+lysis+of+cell+(Yong+et.al.+manuscript+submitted).++In+this+study%2C+we+tested+if+rapid+lysis+by+TDM-specific+hydrolase+(rTDMH)+can+be+exploited+in+conjunction+with+the+RT-PCR+to+develop+a+sensitive+diagnosis+of+tuberculosis.%0D%0A%0D%0AResults+demonstrated+that+by+incubation+of+both+attenuated+M.+tuberculosis%2C+and+virulent+M.+tuberculosis+with+rTDMH+for+lysis+and+subsequent+usage+of+this+lysate+in+a+RT-PCR+assay%2C+yields+sensitive+amplification+of+mycobacterial+DNA.++rTDMH-mediated+lsyis+could+facilitate+amplification+of+even+10+bacilli%2C+the+rTDMH+treated+cells+show+amplification+while+lack+of+treatment+failed+to+detect+these+bacilli++These+results+were+consistent+in+in-vitro+liquid+culture+and+in+complex+sputum+samples+spiked+with+the+mycobacteria%2C+showing+that+incubation+with+rTDMH+can+improve+the+sensitivity+of+the+RT-PCR.+%0D%0A%0D%0AStatement+of+Public+Health+relevance%3A++Using+rTDMH+with+RT-PCR+as+an+improved+diagnostic+tool+for+tuberculosis+due+to+the+rapid%2C+accurate+and+sensitive+nature+of+the+assay+could+provide+the+global+community+with+a+much+better+method+of+diagnosing+a+disease+that+has+plagued+the+world+for+thousands+of+years.+Tuberculosis+infects+9+million+people+and+kills+3+million+people+every+year+and+presently+one-third+of+the+world%E2%80%99s+population+is+infected+with+it.++A+better+diagnostic+tool+could+result+in+reducing+the+spread+of+disease%3B+reducing+the+mortality+associated+with+disease%2C+especially+in+HIV+infected+individuals%3B+and+on+a+broader+scale%2C+could+reduce+the+economic+burden+associated+with+the+disease%0D%0A&rft.date=2012-01-30&rft.type=University+of+Pittsburgh+ETD&rft.type=PeerReviewed&rft.format=application%2Fpdf&rft.language=en&rft.identifier=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F10593%2F1%2FBhatti_aab_etd2011.pdf&rft.identifier=++Bhatti%2C+Alexandra++(2012)+Optimization+of+rTDMH+as+a+Reagent+Toward+Improving+the+Sensitivity+of+the+RT-PCR+Based+Diagnosis+for+Mycobacterium+Tuberculosis.++Master's+Thesis%2C+University+of+Pittsburgh.++++(Unpublished)++