{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,4]],"date-time":"2026-04-04T06:15:23Z","timestamp":1775283323025,"version":"3.50.1"},"reference-count":31,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Clinical Laboratory Analysis"],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>To analyze the clinical diagnostic value for thalassemia genotyping using peripheral blood mRNA.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Total RNA was extracted from 103 pre\u2010genotyped peripheral blood samples, followed by cDNA synthesis. These samples comprised 21 wild\u2010type cases, 49 \u03b1\u2010globin gene mutant cases and 33\n                      <jats:italic>HBB<\/jats:italic>\n                      mutant cases. An Integrated strategy was adopted, combining single\u2010reaction Sanger sequencing with the multiple quantitative fluorescence PCR (MQF\u2010PCR), to analyze the full\u2010length coding sequences and expression level of\n                      <jats:italic>HBA2<\/jats:italic>\n                      \/\n                      <jats:italic>HBA1<\/jats:italic>\n                      \/\n                      <jats:italic>HBB<\/jats:italic>\n                      respectively.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      (1) The substitution mutations located in the coding region (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200914) were successfully detected, and the mutations located in the\n                      <jats:italic>HBB<\/jats:italic>\n                      non\u2010coding region displayed negative results (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200913). The samples with \u03b2\n                      <jats:sup>41\u201342M<\/jats:sup>\n                      \/\u03b2\n                      <jats:sup>N<\/jats:sup>\n                      (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200910) and \u03b2\n                      <jats:sup>71\u201372M<\/jats:sup>\n                      \/\u03b2\n                      <jats:sup>N<\/jats:sup>\n                      (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20094) were not observed to have their small deletion or insertion mutation initially, but they were eventually identified after using allele\u2010specific MQF\u2010PCR. Moreover, MQF\u2010PCR results indicated that the level ranges of Peak area ratio 1 (PAR1, PA\n                      <jats:sub>HBA2<\/jats:sub>\n                      \/PA\n                      <jats:sub>HBA1<\/jats:sub>\n                      ) and Peak area ratio 2 (PAR2, PA\n                      <jats:sub>HBB<\/jats:sub>\n                      \/PA\n                      <jats:sub>HBA<\/jats:sub>\n                      ) can be used to identify the common \u03b1\u2010globin gross\u2010deletion types and to predict mutations in the\n                      <jats:italic>HBB<\/jats:italic>\n                      non\u2010coding region.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      This mRNA\u2010based strategy enabled detection of coding\u2010region mutations in the\n                      <jats:italic>HBA2\/HBA1\/HBB<\/jats:italic>\n                      and prediction of non\u2010coding\u2010region mutations as well as gross deletions with high coverage, low cost and simple operation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/jcla.70186","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T09:43:37Z","timestamp":1772963017000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An Integrated Genotyping Strategy for \u03b1\/\u03b2\u2010Thalassemia: Based on the Analysis of the Coding Sequences and Expression Levels of\n                    <scp>\n                      <i>HBA2<\/i>\n                    <\/scp>\n                    ,\n                    <scp>\n                      <i>HBA1<\/i>\n                    <\/scp>\n                    , and\n                    <scp>\n                      <i>HBB<\/i>\n                    <\/scp>\n                    in Peripheral Blood\n                    <scp>mRNA<\/scp>"],"prefix":"10.1002","volume":"40","author":[{"given":"Hongjian","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Medical Genetics and Prenatal Diagnosis Haikou Maternal and Child Health Hospital  Haikou Hainan China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qi","family":"Yao","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics and Prenatal Diagnosis Haikou Maternal and Child Health Hospital  Haikou Hainan China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mianai","family":"Fu","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics and Prenatal Diagnosis Haikou Maternal and Child Health Hospital  Haikou Hainan China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,2,24]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.clinbiochem.2019.06.005"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2020.00074"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.3389\/fgene.2022.1001369"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0089855"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcmd.2014.01.003"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.3389\/fgene.2023.1126099"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1186\/1750\u20101172\u20105\u201011"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/gim.2016.173"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.12527"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1177\/03000605221078785"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12863\u2010017\u20100594\u20103"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmoldx.2020.03.002"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0102243"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmoldx.2020.05.004"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2017.08.015"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/fped.2022.1015769"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.3390\/diagnostics13030373"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1080\/16078454.2024.2339559"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21186671"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/mgg3.1835"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1136\/jcp.2004.025296"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkae1090"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00439\u2010020\u201002199\u20103"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2890"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.3389\/fped.2023.1139387"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1186\/1750\u20101172\u20105\u201013"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcla.25104"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmoldx.2011.03.005"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12881\u2010017\u20100428\u20101"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcla.21594"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)66871-1"}],"container-title":["Journal 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