{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T17:30:50Z","timestamp":1775064650784,"version":"3.50.1"},"reference-count":17,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2012,10,29]],"date-time":"2012-10-29T00:00:00Z","timestamp":1351468800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion Medicine"],"published-print":{"date-parts":[[2013,2]]},"abstract":"<jats:title>SUMMARY<\/jats:title><jats:sec><jats:title>Objectives\/Aims<\/jats:title><jats:p>This work aims to develop a multiplex polymerase chain reaction combined with <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> pooling for mass screening for rare blood types.<\/jats:p><\/jats:sec><jats:sec><jats:title>Background<\/jats:title><jats:p>The differences in most blood group antigens are associated with single\u2010nucleotide polymorphisms (<jats:styled-content style=\"fixed-case\">SNPs<\/jats:styled-content>), which are used in detecting blood antigen expression at the molecular level. However, all existing sequence\u2010specific primers polymerase chain reaction (<jats:styled-content style=\"fixed-case\">PCR\u2010SSP<\/jats:styled-content>) assays for blood typing genotype one or several <jats:styled-content style=\"fixed-case\">SNPs<\/jats:styled-content> individually. <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> pooling is a way that reduces the amount of genotyping required.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>A sensitive multiplex <jats:styled-content style=\"fixed-case\">PCR\u2010SSP<\/jats:styled-content> assay testing pooled <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> was established to detect the rare <jats:italic>Fy<jats:sup>b<\/jats:sup><\/jats:italic> and <jats:italic>S<\/jats:italic> alleles. It was applied to screen a total of 4490 donor samples via testing 898 <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> pools. The samples in the positive pools were further tested individually. Then the positive samples, including Fy(a\u2212b+)\/Fy(a+b+) and S+s\u2212\/S+s+ genotypes, were tested via two <jats:styled-content style=\"fixed-case\">PCR\u2010SSP<\/jats:styled-content> assays for alleles <jats:italic>Fy<jats:sup>a<\/jats:sup><\/jats:italic> and <jats:italic>s<\/jats:italic>. The rare genotypes Fy(a\u2212b+) and S+s\u2212 were verified using serologic tests and sequencing analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Two hundred and fifty\u2010four donors were tested positive for the <jats:italic>Fy<jats:sup>b<\/jats:sup><\/jats:italic> allele, whereas 101 donors were positive for the <jats:italic>S<\/jats:italic> allele. Among the 254 Fy(b+) donors, 5 were Fy(a\u2212b+) and 249 were Fy(a+b+). Among the 101 S+ donors, 3 were S+s\u2212 and 98 were S+s+. The rare <jats:italic>Fy<jats:sup>b<\/jats:sup><\/jats:italic> and <jats:italic>S<\/jats:italic> alleles comprised 2\u00b728 and 1\u00b716%, respectively. The <jats:styled-content style=\"fixed-case\">PCR\u2010SSP<\/jats:styled-content> assays were confirmed by sequencing analysis and serological test.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>A multiplex <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> assay was combined with <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> pooling to reduce the number of tests required, making large\u2010scale screening feasible.<\/jats:p><\/jats:sec>","DOI":"10.1111\/j.1365-3148.2012.01198.x","type":"journal-article","created":{"date-parts":[[2012,10,30]],"date-time":"2012-10-30T01:36:41Z","timestamp":1351561001000},"page":"42-47","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Multiplex polymerase chain reaction with <scp>DNA<\/scp> pooling: a cost\u2010effective strategy of genotyping rare blood 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