Microbiological applications of high-resolution melting analysis

Abstract

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.

Fig 1

High-resolution melting curves for six internal fragments from the multilocus sequence typing loci of Staphylococcus aureus. The number of G+C residues contained in the fragment labels the curves. For example, in panel A, the melting curve variants of a 181-bp internal fragment of arcC demonstrate an increasing melting temperature (T_m) as the number of G+C residues increases from 51 to 54. In panel B, there is evidence of two melting domains in this 140-bp internal fragment of aroE, resulting in the ability to discriminate variants of the same %G+C. These additional curves have been labeled 23.5 and 24.5. In panel E, melting curves from variants with 64 and 65 G+C residues could not be consistently discriminated for this large 219-bp internal fragment of tpi and have been labeled 64/65. The other fragments are from gmk (C), pta (D), and tpi (F) (a region of tpi different from that represented in panel E). (Reproduced from reference .).