BACKGROUND Single-base pair missense mutations in exons 10, 11, 13, 14, 15, and 16 of the RET protooncogene are associated with the autosomal dominant multiple endocrine neoplasia type 2 (MEN2) syndromes: MEN2A, MEN2B, and familial medullary thyroid carcinoma. The current widely used approach for RET mutation detection is sequencing of the exons. METHODS Because RET mutations are rare and the...

High-resolution melting of polymerase chain reaction (PCR) products can detect heterozygous mutations and most homozygous mutations without electrophoretic or chromatographic separations. However, some homozygous single nucleotide polymorphism (SNPs) have melting curves identical to that of the wild-type, as predicted by nearest neighbor thermodynamic models. In these cases, if DNA of a known r...

Single nucleotide polymorphisms (SNPs) are single base pair mutations that provide new approaches to studies of allele transcript abundances. High-resolution DNA melting curve (HRM) analysis using a LightScanner (Hi-Res Melting system with Idaho's LC Green) provides post-PCR detection of mutations and SNPs in genomic DNA. This study investigated whether the HRM analysis can distinguish alleles ...

BACKGROUND Microsatellite instability (MSI) is an important marker for screening for hereditary nonpolyposis colorectal cancer (Lynch syndrome) as well as a prognostic and predictive marker for sporadic colorectal cancer (CRC). The mononucleotide microsatellite marker panel is a well-established and superior alternative to the traditional Bethesda MSI analysis panel, and does not require testin...

HIV viruses are usually genetically homogeneous shortly after infection, and become more heterogeneous over time. We developed a high-resolution melting (HRM) assay to analyze HIV diversity without sequencing. Plasma samples from the HIVNET 012 trial were obtained from nine Ugandan mother-infant pairs. DNA amplified from the HIV gag region was analyzed to determine the number of degrees over wh...

Methylation of cytosine in DNA is an epigenetic mark that is important for genome stability, transcriptional regulation of endogenous genes, and permanent silencing of transposable elements and viral sequences. Methylcytosine, sometimes referred to as the 5th base of DNA, constitutes 3%–6% of all cytosines in the human genome and occurs almost exclusively within the context of CpG dinucleotides...

Infections caused by non-tuberculous mycobacteria (NTM) is increasing wordwide. Due to the difference in treatment of NTM infections and tuberculosis, rapid species identification of mycobacterial clinical isolates is necessary for the effective management of mycobacterial diseases treatment and their control strategy. In this study, a cost-effective technique, real-time PCR coupled with high-r...

25 A single-tube method – ligation mediated real-time PCR high-resolution melt analysis 26 (LMqPCR HRMA) – was modified for rapid typing of ESKAPE-pathogens. A 97 % 27 agreement (60/62 isolates) was achieved in comparison to PFGE which indicates that 28 LMqPCR HRMA is a rapid and accurate screening tool to monitor for nosocomial outbreaks. 29 on O cber 1, 2017 by gest ht://jcm .sm .rg/ D ow nla...