{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,10]],"date-time":"2025-11-10T13:49:50Z","timestamp":1762782590966,"version":"3.40.5"},"reference-count":37,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2021,6,14]],"date-time":"2021-06-14T00:00:00Z","timestamp":1623628800000},"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":["Int J Lab Hematology"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p><jats:italic>Calreticulin<\/jats:italic> (<jats:italic>CALR<\/jats:italic>) mutations in myeloproliferative neoplasms (MPN) have been reported to be key markers in the molecular diagnosis, particularly in patients lacking <jats:italic>JAK2<\/jats:italic> V617F mutation. In most current reports, <jats:italic>CALR<\/jats:italic> mutations were analysed by either allele\u2010specific PCR (AS\u2010PCR), or the more expensive quantitative real\u2010time PCR, pyrosequencing and next\u2010generation sequencing. Hence, we report the use of an alternative method, the conformation sensitive gel electrophoresis (CSGE) for the detection of <jats:italic>CALR<\/jats:italic> mutations in <jats:italic>BCR\u2010ABL1<\/jats:italic>\u2010negative MPN patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Forty <jats:italic>BCR\u2010ABL1<\/jats:italic>\u2010negative MPN patients\u2019 DNA: 19 polycythemia vera (PV), 7 essential thrombocytosis (ET) and 14 primary myelofibrosis (PMF), were screened for <jats:italic>CALR<\/jats:italic> mutations by CSGE. PCR primers were designed to amplify sequences spanning between exons 8 and 9 to target the mutation hotspots in <jats:italic>CALR<\/jats:italic>. Amplicons displaying abnormal CSGE profiles by electrophoresis were directly sequenced, and results were analysed by BioEdit Sequence Alignment Editor v7.2.6. CSGE results were compared with AS\u2010PCR and confirmed by Sanger sequencing.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>CSGE identified 4 types of mutations; 2 PMF patients with either <jats:italic>CALR<\/jats:italic> type 1 (c.1099_1150del52) or type 2 (c.1155_1156insTTGTC), 1 ET patient with nucleotide deletion (c.1121delA) and insertion (c.1190insA) and 1 PV patient with p.K368del (c.1102_1104delAAG) and insertion (c.1135insA) inframe mutations. Three patients have an altered KDEL motif at the C\u2010terminal of CALR protein. In comparison, AS\u2010PCR only able to detect two PMF patients with mutations, either type 1 and type 2.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>CSGE is inexpensive, sensitive and reliable alternative method for the detection of <jats:italic>CALR<\/jats:italic> mutations in <jats:italic>BCR\u2010ABL1<\/jats:italic>\u2010negative MPN patients.<\/jats:p><\/jats:sec>","DOI":"10.1111\/ijlh.13628","type":"journal-article","created":{"date-parts":[[2021,6,14]],"date-time":"2021-06-14T16:06:05Z","timestamp":1623686765000},"page":"1451-1457","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Conformation sensitive gel electrophoresis for the detection of calreticulin mutations in BCR\u2010ABL1\u2010negative myeloproliferative neoplasms"],"prefix":"10.1111","volume":"43","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1772-3515","authenticated-orcid":false,"given":"Nur Atikah","family":"Zakaria","sequence":"first","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Norfifiana Alisa","family":"Rosle","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3827-7112","authenticated-orcid":false,"given":"Mat Jusoh","family":"Siti Asmaa","sequence":"additional","affiliation":[{"name":"School of Health Sciences Universiti Sains Malaysia Kelantan Malaysia"},{"name":"Universiti Sains Malaysia (USM)\u2010RIKEN Interdisciplinary Collaboration for Advanced Sciences (URICAS) Penang Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1581-8254","authenticated-orcid":false,"given":"Sudin","family":"Aziee","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4434-166X","authenticated-orcid":false,"given":"Mohd Yassim","family":"Haiyuni","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nurul Ameera","family":"Samat","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0899-0443","authenticated-orcid":false,"given":"Azlan","family":"Husin","sequence":"additional","affiliation":[{"name":"Department of Medicine School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"},{"name":"Hospital USM Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0493-1390","authenticated-orcid":false,"given":"Rosline","family":"Hassan","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"},{"name":"Hospital USM Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9104-7511","authenticated-orcid":false,"given":"Marini","family":"Ramli","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"},{"name":"Hospital USM Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8950-5715","authenticated-orcid":false,"given":"Shafini","family":"Mohamed Yusoff","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"},{"name":"Hospital USM Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ibrahim Khidir","family":"Ibrahim","sequence":"additional","affiliation":[{"name":"Faculty of Medical Laboratory Sciences Department of Haematology Al\u2010Neelain University Khartoum Sudan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6194-2729","authenticated-orcid":false,"given":"Hamid Ali Nagi","family":"Al\u2010Jamal","sequence":"additional","affiliation":[{"name":"Diagnostic and Biomedicine Faculty of Health Sciences Universiti Sultan Zainal Abidin Terengganu Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6344-0220","authenticated-orcid":false,"given":"Muhammad Farid","family":"Johan","sequence":"additional","affiliation":[{"name":"Department of Haematology School of Medical Sciences Universiti Sains Malaysia Kelantan Malaysia"},{"name":"Hospital USM Universiti Sains Malaysia Kelantan Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,6,14]]},"reference":[{"volume-title":"\"Essential Hematology.\" Essential Hematology","year":"2011","author":"Hoffbrand AV","key":"e_1_2_10_2_1"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00277-007-0403-6"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(05)71142-9"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1311347"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/leu.2013.382"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1312542"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2016-03-643544"},{"key":"e_1_2_10_9_1","first-page":"16","article-title":"Calreticulin (CALR) mutation in myeloproliferative neoplasms (MPNs)","volume":"2","author":"Luo W","year":"2015","journal-title":"Stem Cell Investig"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.3390\/membranes4030630"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.09-145482"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20081847"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2014-03-530865"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V124.21.157.157"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12185-017-2246-9"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.3343\/alm.2016.36.4.291"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-56236-x"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cca.2018.05.001"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0227986"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.13282"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.25307"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60761-947-5_2"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.7314\/APJCP.2015.16.12.4869"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.21.10325"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1182\/asheducation-2016.1.534"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1309\/AJCPUAAC16LIWZMM"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.4.1681"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.3343\/alm.2015.35.1.22"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2012-05-431791"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.leu.2404730"},{"issue":"1","key":"e_1_2_10_31_1","first-page":"34","article-title":"The concomitant occurrence of JAK2V617F mutation and BCR\/ABL transcript with phenotypic expression \u2013an overlapping myeloproliferative disorder or two distinct diseases?\u2013case report","volume":"6","author":"Ursuleac I","year":"2013","journal-title":"J Med Life"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1177\/1087057104266390"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1038\/leu.2015.277"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1007\/s004399900092"},{"key":"e_1_2_10_35_1","first-page":"1129","article-title":"Calreticulin gene exon 9 frameshift mutations in patients with thrombocytosis","volume":"28","author":"Tefferi A","year":"2014","journal-title":"Leukemia"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.14113"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.4274\/tjh.2015.0220"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2014-05-578435"}],"container-title":["International Journal of Laboratory Hematology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/ijlh.13628","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/ijlh.13628","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/ijlh.13628","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T08:54:35Z","timestamp":1693212875000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/ijlh.13628"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,14]]},"references-count":37,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2021,12]]}},"alternative-id":["10.1111\/ijlh.13628"],"URL":"https:\/\/doi.org\/10.1111\/ijlh.13628","archive":["Portico"],"relation":{},"ISSN":["1751-5521","1751-553X"],"issn-type":[{"type":"print","value":"1751-5521"},{"type":"electronic","value":"1751-553X"}],"subject":[],"published":{"date-parts":[[2021,6,14]]},"assertion":[{"value":"2021-01-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-05-25","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-06-14","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}