{"id":2415,"date":"2025-07-29T13:44:54","date_gmt":"2025-07-29T11:44:54","guid":{"rendered":"https:\/\/biomol.es\/?p=2415"},"modified":"2025-07-29T13:44:56","modified_gmt":"2025-07-29T11:44:56","slug":"clinical-implementation-of-the-molecular-algorithm-for-ph-negative-mpn-in-collaboration-with-the-hospital-of-jerez","status":"publish","type":"post","link":"https:\/\/biomol.es\/en\/clinical-implementation-of-the-molecular-algorithm-for-ph-negative-mpn-in-collaboration-with-the-hospital-of-jerez\/","title":{"rendered":"Clinical Implementation of the Molecular Algorithm for Ph-Negative MPN in Collaboration with the Hospital of Jerez"},"content":{"rendered":"\n<p><br>From the Support and Applications Department at BIOMOL, we share a recent case of implementing a complete diagnostic workflow for <strong>Philadelphia-negative Myeloproliferative Neoplasms (Ph-negative MPNs)<\/strong>, developed in collaboration with the Hematology Department of the University Hospital of Jerez de la Frontera.<\/p>\n\n\n\n<p>The objective was to structure a <strong>sequential algorithm<\/strong> that enables accurate identification of mutations in <strong>BCR-ABL1, JAK2, CALR, and MPL<\/strong>, integrating technologies such as <strong>real-time PCR, HRM<\/strong>, and <strong>next-generation sequencing (NGS)<\/strong>, while ensuring standardization across <strong>7500 Fast<\/strong> and <strong>SeqStudio<\/strong> platforms.<br><br>Standardization: Equipment and Protocols<\/p>\n\n\n\n<p><strong>1. BCR-ABL1 \u2013 Screening and Quantification<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>For BCR-ABL1 screening and quantification, a transition was made from the <strong>LightCycler (LC)<\/strong> system to the <strong>7500 Fast Real-Time PCR<\/strong> system (Applied Biosystems).<\/li>\n\n\n\n<li>This migration was carried out without requiring additional modifications, maintaining the previous detection standards.<\/li>\n\n\n\n<li>A specific standard curve for the <strong>p190 isoform<\/strong> was developed, enabling robust quantification.<\/li>\n\n\n\n<li>A curve for the <strong>p210 isoform<\/strong> was also validated, although it continues to be commonly quantified using an alternative methodology, as per the laboratory\u2019s decision.<\/li>\n<\/ul>\n\n\n\n<p><strong>2. JAK2 V617F<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>The diagnosis of this mutation, present in <strong>PV, ET, and PMF<\/strong>, continues to be performed using client-specific probes and primers already optimized for the 7500 system. No intervention from BIOMOL was required.<\/li>\n<\/ul>\n\n\n\n<p><strong>3. JAK2 Exon 12 \u2013 HRM<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>The protocol for <strong>High-Resolution Melting (HRM)<\/strong> analysis was reinstalled on the 7500 platform, particularly useful in suspected <strong>Polycythemia Vera<\/strong> cases with negative JAK2 V617F.<\/li>\n\n\n\n<li>This approach allows discrimination of less common variants not detected by ASO-PCR.<\/li>\n<\/ul>\n\n\n\n<p><strong>4. CALR \u2013 Fragment Analysis<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>The <strong>CALR assay<\/strong> is performed via fragment analysis on the <strong>SeqStudio system<\/strong>.<\/li>\n\n\n\n<li>Active technical support was provided due to challenges with visualization and analysis. After adjusting electrophoresis parameters and fragment size, interpretable and reproducible profiles were achieved.<\/li>\n<\/ul>\n\n\n\n<p><\/p>\n\n\n\n<p>Clinical Results and Validation<\/p>\n\n\n\n<p>The algorithm was applied to <strong>414 patients<\/strong> with suspected Ph-negative MPN:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>BCR-ABL1<\/strong> was negative in all cases.<\/li>\n\n\n\n<li><strong>40 patients<\/strong> were identified as <strong>JAK2 V617F positive<\/strong>.<\/li>\n\n\n\n<li>For <strong>JAK2-negative<\/strong> patients, <strong>Exon 12<\/strong> was analyzed, followed by <strong>CALR and MPL<\/strong> sequencing.<br>In a sub-cohort negative for both BCR-ABL1 and JAK2, <strong>NGS<\/strong> was applied to optimize the detection of non-canonical mutations.<\/li>\n\n\n\n<li>All positive results for <strong>JAK2, CALR, or MPL<\/strong> were consistent with the final clinical diagnosis, supporting the robustness of the proposed algorithm.<\/li>\n<\/ul>\n\n\n\n<p><\/p>\n\n\n\n<p>Conclusions<\/p>\n\n\n\n<p>The collaboration with the University Hospital of Jerez highlights the importance of specialized technical support in the implementation of complex diagnostic workflows in clinical settings.<\/p>\n\n\n\n<p>BIOMOL\u2019s contribution was crucial in the <strong>standardization of protocols<\/strong>, <strong>adaptation of platforms (7500 Fast and SeqStudio)<\/strong>, and <strong>integration of advanced methodologies such as HRM and NGS<\/strong>. This structured approach ensured <strong>analytical traceability<\/strong> and strengthened the reliability of molecular diagnostics in patients with suspected myeloproliferative neoplasms.<\/p>\n\n\n\n<p>This case reaffirms the value of a well-designed implementation aligned with laboratory practices, and reinforces our commitment to a more <strong>agile, traceable, and effective precision medicine<\/strong>.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>From the Support and Applications Department at BIOMOL, we share a recent case of implementing a complete diagnostic workflow for Philadelphia-negative Myeloproliferative Neoplasms (Ph-negative MPNs), developed in collaboration with the Hematology Department of the University Hospital of Jerez de la Frontera. The objective was to structure a sequential algorithm that enables accurate identification of mutations &hellip;<\/p>\n","protected":false},"author":12,"featured_media":2371,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"inline_featured_image":false,"footnotes":""},"categories":[36,48,49],"tags":[],"class_list":["post-2415","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-biomedicine","category-fragmentation","category-ngs-2"],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.4 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Clinical Implementation of the Molecular Algorithm for Ph-Negative MPN in Collaboration with the Hospital of Jerez - Biomol<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/biomol.es\/en\/clinical-implementation-of-the-molecular-algorithm-for-ph-negative-mpn-in-collaboration-with-the-hospital-of-jerez\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Clinical Implementation of the Molecular Algorithm for Ph-Negative MPN in Collaboration with the Hospital of Jerez - Biomol\" \/>\n<meta property=\"og:description\" content=\"From the Support and Applications Department at BIOMOL, we share a recent case of implementing a complete diagnostic workflow for Philadelphia-negative Myeloproliferative Neoplasms (Ph-negative MPNs), developed in collaboration with the Hematology Department of the University Hospital of Jerez de la Frontera. 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