High‑resolution melting analysis for rapid and sensitive MYD88 screening in chronic lymphocytic leukemia

Abstract

High resolution melting (HRM) assay is a novel technology for the fast, high-throughput, sensitive, post-PCR analysis of genetic mutations. Myeloid differentiation primary response 88 (MYD88) mutations are frequently reported in chronic lymphocytic leukemia (CLL) and confer a worse prognosis. The objective of the present study was to assess the value of HRM analysis for the rapid screening of MYD88 mutations in patients with CLL. Genomic DNA samples were extracted from the bone marrow of 129 newly diagnosed patients with CLL. A plasmid with an MYD88-L265P mutation was constructed, and the p.L265P substitution, which is the predominant MYD88 mutation in CLL, was detected using HRM analysis and direct sequencing. The plasmid pCMV-MYD88-L265P-Mu was successfully constructed as a positive control, and was verified by direct sequencing. The normalized and shifted melting curves of 6/129 (4.65%) samples were clearly different from those of other patients by HRM analysis. In addition, the 794T>C mutation in MYD88 was identified in 6 (4.65%) patients by direct sequencing. Sensitivity evaluation revealed that the HRM assay had a higher sensitivity (to 1% dilution) than direct sequencing, in addition to being convenient and time-saving. The MYD88 p.L256P mutation has been implicated to be associated with adverse prognosis in CLL. HRM analysis has the potential to be a routine prescreening technique to identify the MYD88 p.L256P mutation and may facilitate the clinical treatment of CLL.

Highly Sensitive 3D‐Nanoplasmonic‐Based Epidermal Growth Factor Receptor Mutation Multiplex Assay Chip for Liquid Biopsy

“…Although direct sequencing and NGS detect various mutations, their sensitivity is notably low, typically in the range of 1%-10%. [11,29] Digital PCR, a recently developed method with enhanced sensitivity (0.01%), requires numerous primers for the detection of various mutations. [11,30] In contrast, our assay method showed remarkable sensitivity of 1 Â 10 À9 %, surpassing that of digital PCR by 10 7 -fold.…”

MYD88 in the driver’s seat of B-cell lymphomagenesis: from molecular mechanisms to clinical implications

“…83 Furthermore, MYD88(L265P) is absent in primary mediastinal large Bcell lymphoma 2,3,94 and primary cutaneous follicle center lymphoma, [71][72][73] and rarely present in hairy cell leukemia (1.1%), 22,30,[57][58][59] plasma cell myeloma (1.5%), 18,22,23,43,106,107 Burkitt lymphoma (1.5%), 2,74 follicular lymphoma (1.9%), 18,22,23,67,68 and CLL (2.5%). 18,[22][23][24]28,[38][39][40][41][42][43][44][45][46][47][48][49][50][51][52]…”

Feasibility of melting fingerprint obtained from ISSR-HRM curves for marine mammal species identification

“…This approach has several notable advantages such as reducing the chances of cross-contamination and the fact that it does not require the handling of hazardous materials ( Erdem et al, 2016 ; Gopaul et al, 2014 ). Additionally, it is recognized as being less time-consuming ( Jiang et al, 2019 ). Besides, specimens could be distinguished by graph changes in the melting curves that are generated by HRM and are easily visualized ( Jin et al, 2014 ).…”