BACKGROUND Genotyping of human papillomarvirus (HPV) is crucial for patient management in a clinical setting. This study accesses the combined use of broad-range real-time PCR and high-resolution melting (HRM) analysis for rapid identification of HPV genotypes. METHODS Genomic DNA sequences of 8 high-risk genotypes (HPV16/18/39/45/52/56/58/68) were subject to bioinformatic analysis to select ...

High resolution melting (HRM) is a novel, closed-tube, post-PCR technique allowing genomic researchers to easily analyze genetic variations in PCR amplicons. This technical note describes general steps of setting up HRM-based PCR assays, with a special focus on ways how to optimize procedures for gene scanning experiments. The novel High Resolution Melting (HRM) technique enables researchers to...

DNA methylation is a key epigenetic mechanism that regulates gene expression and chromatin structure. This application note describes a workflow for analysis of DNA methylation in specific loci by methylationsensitive high-resolution melting (MS-HRM) followed by DNA sequencing of the reaction products (Figure 1). The workflow relies on Applied Biosystems® real-time PCR and capillary electrophor...

Pasteurellaceae are among the most prevalent bacterial pathogens isolated from mice housed in experimental animal facilities. Reliable detection and differentiation of Pasteurellaceae are essential for high-quality health monitoring. In this study, we combined a real-time PCR assay amplifying a variable region in the 16S rRNA sequence with high-resolution melting curve analysis (HRM) to identif...

High-resolution melting (HRM) analysis uses real-time PCR instrumentation to interrogate DNA sequence variation and is a low-cost, single-step, closed-tube method. Here we describe HRM technology and provide examples of varied clinical microbiological applications to highlight the strengths and limitations of HRM analysis.

BACKGROUND Prevention and control of thalassemia requires simple, rapid, and accurate screening tests for carrier couples who are at risk of conceiving fetuses with severe thalassemia. METHODS Single-tube multiplex real-time PCR with SYBR Green1 and high-resolution melting (HRM) analysis were used for the identification of α-thalassemia-1 Southeast Asian (SEA) and Thai type deletions and β-th...

We developed a simple, rapid, high-throughput detection and genotyping method for noroviruses using real-time reverse transcription-PCR (RT-PCR) and high-resolution melting (HRM) analysis to create a difference plot. The capsid gene was amplified by real-time RT-PCR in the presence of ResoLight HRM dye, a saturating DNA dye. Following optimization of the HRM assay conditions, the major noroviru...

Objective(s): Rapid and accurate detection of Brucella abortus and Brucella melitensis from clinical samples is so important because antibiotic treatment has major side effects. This study reveals a new method in detection of clinical samples of brucellosis using real-time PCR and high-resolution melt (HRM) curve analysis. Materials and Methods: 160 brucellosis suspicious samples with more tha...