ABSTRACT
CXCR4 mutations impact disease presentation and treatment outcomes in Waldenström macroglobulinemia. Current techniques used for CXCR4 mutation detection have a number of limitations. The aim of the present study was to develop and analytically validate a novel droplet digital PCR (ddPCR) assay for the simultaneous detection of five of the most common CXCR4 mutations in bone marrow (BM). In silico novel primers and probes designed for simultaneous detection of five hotspot mutations of CXCR4 were first performed. Experimental conditions were optimized, and the assay was analytically validated. The developed assay was further applied in 95 BM samples from patients with IgM gammopathy, 7 BM samples from patients with non-IgM gammopathy and 12 PBMCs from healthy donors, whereas a direct comparison study of Sanger sequencing and allele-specific PCR was performed by using 95 and 39 identical patient tumor DNA samples, respectively. The drop-off ddPCR assay is a robust, cost-effective, highly sensitive, and highly specific screening tool for CXCR4 mutations. Of 95 patients with IgM gammopathy samples, 27 had at least one CXCR4 mutation in their BM samples. With Sanger sequencing, 12 of the 95 samples tested positive, whereas the direct comparison of the developed assay with allele-specific PCR revealed substantial agreement. The clinical performance of the developed assay will be prospectively evaluated in a large number of patients, and the applicability of this assay will be further evaluated.
Subject(s)
Lymphoma, B-Cell , Waldenstrom Macroglobulinemia , Humans , Waldenstrom Macroglobulinemia/diagnosis , Waldenstrom Macroglobulinemia/genetics , Mutation , Polymerase Chain Reaction/methods , Alleles , Lymphoma, B-Cell/genetics , Receptors, CXCR4/geneticsABSTRACT
To investigate the implementation of home blood pressure monitoring (HBPM) guidelines, a phone survey was performed in 366 primary care physicians (PCPs). Of the PCPs, 90% routinely used HBPM for white-coat hypertension, treatment titration, and diagnosis. Thirty percent trusted HBPM more than office measurements. Reported drawbacks were questionable reliability of patients' reports and devices inaccuracy. Thirty-one percent advised patients on device selection, 38% were aware of validated devices, and 69% reviewed (not averaged) the readings. Seventy-nine percent used higher than recommended threshold for hypertension diagnosis. Although PCPs routinely use HBPM, there are important gaps in their knowledge and educational activities are required.
