Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan.
Department of Laboratory Medicine, Nagasaki University Hospital, Nagasaki, Japan; Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Clin Chim Acta. 2016 Nov 1;462:166-173. doi: 10.1016/j.cca.2016.09.023. Epub 2016 Sep 29.
Recently, novel calreticulin (CALR) mutations were discovered in Janus kinase 2 (JAK2) non-mutated myelofibrosis (PMF) and essential thrombocythemia (ET) cases, with a frequency of 60-80%. We examined clinical correlations and CALR mutation frequency in our myeloproliferative neoplasms (MPN) cases, and introduce an effective test method for use in clinical practice.
We examined 177 samples previously investigated for the JAK2 mutation for differential diagnosis of MPN. JAK2 and CALR mutations were analyzed using melting curve analysis and microchip electrophoresis, respectively. Next, we constructed a test for simultaneous screening of the JAK2 and CALR mutations utilizing high resolution melting (HRM).
Among 99 MPN cases, 60 possessed the JAK2 mutation alone. Of the 39 MPN cases without the JAK2 mutation, 14 were positive for the CALR mutation, all of which were ET. Using our novel screening test for the JAK2 and CALR mutations by HRM, the concordance rate of conventional analysis with HRM was 96% for the JAK2 mutation and 95% for the CALR mutation.
Our novel simultaneous screening test for the JAK2 and CALR gene mutations with HRM is useful for diagnosis of MPN.
最近，在 Janus 激酶 2（JAK2）未突变的骨髓纤维化（PMF）和原发性血小板增多症（ET）病例中发现了新型钙网蛋白（CALR）突变，其频率为 60-80%。我们研究了CALR 突变在我们的骨髓增殖性肿瘤（MPN）病例中的临床相关性和突变频率，并介绍了一种在临床实践中有效的测试方法。
我们检查了 177 个先前用于 JAK2 突变的样本，用于 MPN 的鉴别诊断。使用熔解曲线分析和微芯片电泳分别分析 JAK2 和 CALR 突变。接下来，我们构建了一种利用高分辨率熔解（HRM）同时筛查 JAK2 和 CALR 突变的测试。
在 99 例 MPN 病例中，60 例单独存在 JAK2 突变。在 39 例无 JAK2 突变的 MPN 病例中，14 例存在 CALR 突变，均为 ET。使用我们通过 HRM 对 JAK2 和 CALR 突变进行的新型筛选测试，传统分析与 HRM 的一致性率为 JAK2 突变 96%，CALR 突变 95%。
我们通过 HRM 对 JAK2 和 CALR 基因突变进行的新型同时筛选测试对 MPN 的诊断有用。
