?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%2F36401%2F&rft.title=Development+of+a+rapid+clinical+flow+cytometric+assay+of+neutrophil+cell-surface+biomarkers&rft.creator=Weissman%2C+Alexandra&rft.description=Rationale%3A+Neutrophils+are+upregulated+in+infectious+processes.+They+can+be+measured+as+surrogates+for+the+presence+of+an+infection.+Flow+cytometry+is+a+common+tool+that+uses+fluorochrome-conjugated+antibodies+specific+to+cell-surface+biomarkers+to+identify+cells+or+biomarkers+of+interest.+Many+clinical+flow+cytometry+protocols+exist+that+assess+lymphocytes%2C+but+fewer+protocols+exist+for+rapid+clinical+evaluation+of+neutrophils.+Expression+of+neutrophil+cell-surface+markers+integrin+%CE%B19%CE%B21+and+CD11b+has+been+shown+to+increase+in+elderly+patients+with+bacterial+pneumonia.+A+rapid+clinical+test+for+these+neutrophil+cell-surface+markers+could+help+to+expedite+appropriate+antibiotic+therapy+to+patients+suffering+from+complex+critical+illnesses+complicated+by+bacterial+pneumonia+for+which+other+diagnostic+methods+are+rendered+inadequate.%0D%0AMethods%3A+We+developed+a+rapid+lyse%2C+no-wash+clinical+protocol+for+flow+cytometric+analysis+of+neutrophil+cell-surface+biomarkers+integrin+%CE%B19%CE%B21+and+CD11b+from+whole+blood+samples.+Assay+development+included+selecting+appropriate+neutrophil-specific+biomarkers+and+their+corresponding+fluorochrome+profile%2C+determining+the+most+consistent+blood+acquisition+method+(BAM)+for+obtaining+samples+for+analysis%2C+and+evaluating+the+effect+of+preservative+type+and+time+on+neutrophil+absolute+count+(ANC)%2C+integrin+%CE%B19%CE%B21+mean+fluorescence+intensity+(MFI)%2C+and+CD11b+MFI.%0D%0AResults%3A+There+were+significant+differences+in+ANC%2C+CD11b+MFI%2C+and+integrin+%CE%B19%CE%B21+MFI+over+time+for+each+BAM+and+Preservative+type.+ANC+demonstrated+the+least+variability+in+Citrate%2C+while+CD11b+MFI+and+integrin+%CE%B19%CE%B21+MFI+were+most+consistent+in+Heparin.+Therefore%2C+Heparin+was+selected+as+the+BAM+of+choice.+Streck+preservative+solution+and+Streck+Blood+Collection+Tube+(BCT)+demonstrated+the+least+variability+in+ANC%2C+integrin+%CE%B19%CE%B21+MFI%2C+and+CD11b+MFI.%0D%0AConclusions%3A+Pneumonia+is+difficult+to+detect+in+patients+with+critical+illness%2C+where+the+usual+diagnostic+tools+are+rendered+inadequate+secondary+to+multi-system+organ+failure.+More+accurate+methods+of+diagnosis+for+pneumonia+can+enhance+targeted+antibiotic+therapy+and+are+of+paramount+public+health+importance+to+improve+patient+care.+Development+of+a+rapid+clinical+flow+cytometric+protocol+is+the+first+step+toward+precision+methods+of+pneumonia+diagnosis%3B+therefore%2C+a+rapid+lyse%2Fno-wash+clinical+flow+cytometric+assay+for+whole+blood+specimens+was+developed+for+identification+of+neutrophil+cell-surface+biomarkers+CD11b+and+integrin+%CE%B19%CE%B21.+Heparin+was+the+least+variable+BAM+for+assessing+CD11b+MFI+and+integrin+%CE%B19%CE%B21+MFI%2C+while+Streck+preservative+solution+and+Streck+BCT+demonstrated+the+least+variability+over+24+hours+from+specimen+collection.+Heparin+is+a+common+type+of+venipuncture+tube+in+hospitals%2C+it+is+not+as+cost+effective+or+feasible+to+purchase+a+separate+blood+collection+tube+(e.g.+Streck+BCT)+for+a+single+laboratory+test.+Given+these+real-world+concerns%2C+Streck+solution+added+to+samples+collected+in+heparin+venipuncture+tubes+is+the+most+reasonable+method+by+which+to+perform+the+rapid+lyse%2Fno-wash+neutrophil+flow+cytometric+assay.&rft.date=2019-06-19&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%2F36401%2F1%2FWeissmanAlexandra_%2520MPHThesis_4_2019.pdf&rft.identifier=++Weissman%2C+Alexandra++(2019)+Development+of+a+rapid+clinical+flow+cytometric+assay+of+neutrophil+cell-surface+biomarkers.++Master's+Thesis%2C+University+of+Pittsburgh.++++(Unpublished)++