{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T06:51:43Z","timestamp":1791096703490,"version":"4.1.0"},"reference-count":15,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2013,2,10]],"date-time":"2013-02-10T00:00:00Z","timestamp":1360454400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Tissue Antigens"],"published-print":{"date-parts":[[2013,3]]},"abstract":"Abstract<\/jats:title>The human leukocyte antigen (HLA<\/jats:styled-content>) class I and class II<\/jats:styled-content> loci are the most polymorphic genes in the human genome; distinguishing the thousands of HLA<\/jats:styled-content> alleles is challenging. Next generation sequencing of exonic amplicons with the 454 genome sequence (GS<\/jats:styled-content>) FLX<\/jats:styled-content> System and Conexio<\/jats:sc> Assign<\/jats:sc> ATF 454 software provides high resolution, high throughput HLA<\/jats:styled-content> genotyping for eight class I and class II<\/jats:styled-content> loci. HLA<\/jats:styled-content> typing of potential donors for unrelated bone marrow donor registries typically uses a subset of these loci at high sample throughput and low cost per sample. The Fluidigm Access Array System enables the incorporation of 48 different multiplex identifiers (MIDs<\/jats:styled-content>) corresponding to 48 genomic DNA<\/jats:styled-content> samples with up to 48 different primer pairs in a microfluidic device generating 2304 parallel polymerase chain reactions (PCRs<\/jats:styled-content>). Minimal volumes of reagents are used. During genomic PCR<\/jats:styled-content>, in this 4\u2010primer system, the outer set of primers containing the MID<\/jats:styled-content> and the 454 adaptor sequences are incorporated into an amplicon generated by the inner HLA<\/jats:styled-content> target\u2010specific primers each containing a common sequence tag at the 5\u2032 end of the forward and reverse primers. Pools of the resulting amplicons are used for emulsion PCR<\/jats:styled-content> and clonal sequencing on the 454 Life Sciences GS FLX<\/jats:styled-content> System, followed by genotyping with Conexio<\/jats:sc> software. We have genotyped 192 samples with 100% concordance to known genotypes using 8 primer pairs (covering exons 2 and 3 of HLA<\/jats:styled-content>\u2010A, B and C, and exon 2 of DRB1<\/jats:styled-content>, 3\/4\/5 and DQB1<\/jats:styled-content>) and 96 MIDs<\/jats:styled-content> in a single GS FLX<\/jats:styled-content> run. An average of 166 reads per amplicon was obtained. We have also genotyped 96 samples at high resolution (14 primer pairs covering exons 2, 3, and 4 of the class I loci and exons 2 of DRB1<\/jats:styled-content>, 3\/4\/5, DQA1<\/jats:styled-content>, DQB1<\/jats:styled-content>, DPB1<\/jats:styled-content>, and exon 3 of DQB1<\/jats:styled-content>), recovering an average of 173 sequence reads per amplicon.<\/jats:p>","DOI":"10.1111\/tan.12071","type":"journal-article","created":{"date-parts":[[2013,2,11]],"date-time":"2013-02-11T06:49:40Z","timestamp":1360565380000},"page":"141-149","source":"Crossref","is-referenced-by-count":155,"title":["High throughput HLA<\/scp> genotyping using 454 sequencing and the Fluidigm Access Array\u2122 system for simplified amplicon library preparation"],"prefix":"10.1111","volume":"81","author":[{"given":"P. V.","family":"Moonsamy","sequence":"first","affiliation":[{"name":"Roche Molecular Systems, Inc. Pleasanton CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T.","family":"Williams","sequence":"additional","affiliation":[{"name":"Children's Hospital Oakland Research Institute Oakland CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P.","family":"Bonella","sequence":"additional","affiliation":[{"name":"Children's Hospital Oakland Research Institute Oakland CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. L.","family":"Holcomb","sequence":"additional","affiliation":[{"name":"Roche Molecular Systems, Inc. Pleasanton CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B. N.","family":"H\u00f6glund","sequence":"additional","affiliation":[{"name":"Roche Molecular Systems, Inc. Pleasanton CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Hillman","sequence":"additional","affiliation":[{"name":"Roche Molecular Systems, Inc. Pleasanton CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D.","family":"Goodridge","sequence":"additional","affiliation":[{"name":"Conexio Genomics Perth Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G. S.","family":"Turenchalk","sequence":"additional","affiliation":[{"name":"454 Life Sciences, A Roche Company Branford CT USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L. A.","family":"Blake","sequence":"additional","affiliation":[{"name":"454 Life Sciences, A Roche Company Branford CT USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D. A.","family":"Daigle","sequence":"additional","affiliation":[{"name":"454 Life Sciences, A Roche Company Branford CT USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B. B.","family":"Simen","sequence":"additional","affiliation":[{"name":"454 Life Sciences, A Roche Company Branford CT USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Hamilton","sequence":"additional","affiliation":[{"name":"Fluidigm Corporation South San Francisco CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A. P.","family":"May","sequence":"additional","affiliation":[{"name":"Fluidigm Corporation South San Francisco CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H. A.","family":"Erlich","sequence":"additional","affiliation":[{"name":"Roche Molecular Systems, Inc. Pleasanton CA USA"},{"name":"Children's Hospital Oakland Research Institute Oakland CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2013,2,10]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkq998"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399-0039.2010.01466.x"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399-0039.2009.01345.x"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399-0039.2010.01606.x"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.humimm.2010.06.016"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.humimm.2009.08.009"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2007-06-097386"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2004-03-0803"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.humimm.2011.06.002"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood\u20102011\u201005\u2010353490"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1177\/1740774510373494"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/0198-8859(93)90141-M"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature03959"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.humimm.2012.07.024"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbmt.2008.05.001"}],"container-title":["Tissue Antigens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ftan.12071","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/tan.12071","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,22]],"date-time":"2023-11-22T06:07:14Z","timestamp":1700633234000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/tan.12071"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,2,10]]},"references-count":15,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2013,3]]}},"alternative-id":["10.1111\/tan.12071"],"URL":"https:\/\/doi.org\/10.1111\/tan.12071","archive":["Portico"],"relation":{},"ISSN":["0001-2815","1399-0039"],"issn-type":[{"value":"0001-2815","type":"print"},{"value":"1399-0039","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,2,10]]}}}