?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%2F7042%2F&rft.title=Representational+Difference+Analysis+(RDA)+for+Detection+of+Genetic+Elements+Associated+with+Increased+Incidence+of+Serogroup+C+Neisseria+meningitidis+Infection&rft.creator=Kostelnik%2C+Leah+M.&rft.description=Previous+studies+have+demonstrated+that+the+increased+incidence+of+invasive+disease+caused+by+serogroup+C+Neisseria+meningitidis+in+the+United+States+during+the+1990s+was+attributed+primarily+to+strains+belonging+to+the+ST11+clonal+complex.+Subcapsular+genotyping+of+a+subset+of+isolates+from+Maryland+identified+distinct+%22early%22+and+%22late%22+clones+defined+by+antigenic+shift+at+the+FetA+outer+membrane+protein.+Representational+difference+analysis+(RDA)+was+used+to+identify+additional+genetic+differences+that+may+have+contributed+to+the+emergence+of+the+late+clone.+A+collection+of+serogroup+C+isolates+representative+of+the+early+and+late+clone+was+subjected+to+pulsed+field+gel+electrophoresis+(PFGE)+to+determine+genetic+relatedness+among+the+isolates+and+to+identify+a+candidate+tester%2Fdriver+pair+for+RDA.+RsaI-digested+tester+genomic+DNA+(late+clone)+was+ligated+to+specific+adaptors+followed+by+two+rounds+of+subtractive+hybridization+with+RsaI-digested+driver+genomic+DNA+(early+clone).+PCR+amplification+of+subtracted+tester+DNA+with+adaptor+specific+primers+generated+at+least+three+late+clone-specific+bands+that+were+absent+from+the+early+clone.+These+products+were+cloned+and+sequenced+and+confirmed+by+Southern+blotting+with+tester+and+driver+digoxigenin-labeled+genomic+DNA+probes+to+be+tester+specific.+A+BLAST+search+of+late+clone-specific+sequences+identified+homology+to+either+IS1301+or+pJS-B+plasmid+N.+meningitidis+sequences.+PCR+with+primers+specific+to+either+IS1301+or+pJS-B+plasmid+sequences+amplified+these+elements+from+late+clone+isolates+but+not+from+early+clone+isolates.+Thus%2C+RDA+successfully+identified+two+unique+genetic+elements+present+in+an+emergent+N.+meningitidis+serogroup+C+ST-11+clone+that+had+undergone+antigenic+shift+at+FetA.+Further+investigation+is+required+to+determine+the+potential+role+of+these+elements+in+clonal+emergence+and+N.+meningitidis+pathogenesis.+The+public+health+significance+of+this+project+stems+from+increased+incidence+of+meningococcal+disease+being+a+major+concern%3A+morbidity+and+mortality+increase%2C+outbreaks+produce+panic+and+disruption+in+communities%2C+public+health+agencies+must+respond+for+control+and+prevention%2C+and+mass+immunization+and+antibiotic+prophylaxis+are+often+required.&rft.date=2006-06-07&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%2F7042%2F1%2Fkostelnikleah_edt2006.pdf&rft.identifier=++Kostelnik%2C+Leah+M.++(2006)+Representational+Difference+Analysis+(RDA)+for+Detection+of+Genetic+Elements+Associated+with+Increased+Incidence+of+Serogroup+C+Neisseria+meningitidis+Infection.++Master's+Thesis%2C+University+of+Pittsburgh.++++(Unpublished)++