{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,11,21]],"date-time":"2023-11-21T15:58:53Z","timestamp":1700582333109},"reference-count":12,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2005,6,10]],"date-time":"2005-06-10T00:00:00Z","timestamp":1118361600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Part B Clinical"],"published-print":{"date-parts":[[2005,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Recently, a toxin produced by <jats:italic>Aeromonas hydrophila<\/jats:italic> was demonstrated to bind directly to the glycosyl\u2010phosphatidyl\u2010inositol (GPI) anchor. After coupling it to a fluorescent dye and applying it in fluorescence\u2010activated cell scanning (FACS), this property was exploited to detect GPI\u2010negative cells in the diagnosis of paroxysmal nocturnal haemoglobinuria (PNH).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We used this reagent according to a very simple staining protocol followed by single\u2010colour FACS and compared the results in patients with PNH and normal controls with those obtained with antibody\u2010mediated detection of cells lacking GPI\u2010anchored proteins.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We observed very good concordance between the two methods, with correlation coefficients (R<jats:sup>2<\/jats:sup>) of quantified GPI\u2010deficient cell populations ranging from 0.952 to 0.969. The lower limit of detection was determined at 0.50% GPI\u2010negative cells, which was in the range obtained with double\u2010colour staining with antibodies (0.20\u20131.00%, depending on the antibody). A significant correlation was observed between the fraction of GPI\u2010negative granulocytes and laboratory parameters of haemolysis, with the erythrocyte creatine having the best correlation (R<jats:sup>2<\/jats:sup> = 0.671, <jats:italic>P<\/jats:italic> &lt; 0.0001).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Using this protocol, we were able to reliably diagnose PNH with a high sensitivity. The test allows the identification of GPI\u2010negative granulocyte populations as small as 0.5%. \u00a9 2005 Wiley\u2010Liss, Inc.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.b.20059","type":"journal-article","created":{"date-parts":[[2005,6,10]],"date-time":"2005-06-10T20:46:43Z","timestamp":1118436403000},"page":"13-18","source":"Crossref","is-referenced-by-count":21,"title":["Clinical evaluation of an aerolysin\u2010based screening test for paroxysmal nocturnal haemoglobinuria"],"prefix":"10.1002","volume":"67B","author":[{"given":"Pietro Edmondo","family":"Peghini","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J\u00f6rg","family":"Fehr","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,6,10]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM196607212750308"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V35.4.462.462"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V76.9.1853.1853"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V87.12.5332.bloodjournal87125332"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.1991.tb08060.x"},{"key":"e_1_2_6_7_2","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1128\/jb.163.1.336-340.1985","article-title":"Activation of the hole\u2010forming toxin aerolysin by extracellular processing","volume":"163","author":"Howard SP","year":"1985","journal-title":"J Bacteriol"},{"key":"e_1_2_6_8_2","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1128\/iai.56.5.1249-1253.1988","article-title":"The cytolytic toxin aerolysin must aggregate to disrupt erythrocytes, and aggregation is stimulated by human glycophorin","volume":"56","author":"Garland WJ","year":"1988","journal-title":"Infect Immun"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1159\/000057638"},{"key":"e_1_2_6_10_2","first-page":"472A","article-title":"Diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) and selection of small PNH populations using a novel GPI\u2010anchor binding toxin, aerolysin","volume":"92","author":"Brodsky RA","year":"1998","journal-title":"Blood"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1093\/ajcp\/114.3.459"},{"key":"e_1_2_6_12_2","doi-asserted-by":"crossref","first-page":"966","DOI":"10.1182\/blood.V53.5.966.966","article-title":"Comparison of red\u2010cell creatine level and reticulocyte count in appraising the severity of hemolytic processes","volume":"53","author":"Fehr J","year":"1979","journal-title":"Blood"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.7326\/0003-4819-131-6-199909210-00002"}],"container-title":["Cytometry Part B: Clinical Cytometry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcyto.b.20059","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.b.20059","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,13]],"date-time":"2023-09-13T14:14:45Z","timestamp":1694614485000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.b.20059"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,6,10]]},"references-count":12,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2005,9]]}},"alternative-id":["10.1002\/cyto.b.20059"],"URL":"https:\/\/doi.org\/10.1002\/cyto.b.20059","archive":["Portico"],"relation":{},"ISSN":["1552-4949","1552-4957"],"issn-type":[{"value":"1552-4949","type":"print"},{"value":"1552-4957","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,6,10]]}}}