The Potential of High Resolution Melting Analysis (Hrma) to Streamline, Facilitate and Enrich Routine Diagnostics in Medical Microbiology

Abstract

Background. Routine medical microbiology diagnostics relies on conventional cultivation followed by phenotypic techniques for identification of pathogenic bacteria and fungi. This is not only due to tradition and economy but also because it provides pure culture needed for antibiotic susceptibility testing. This review focuses on the potential of High Resolution Melting Analysis (HRMA) of double-stranded DNA for future routine medical microbiology.Methods and Results. Search of MEDLINE database for publications showing the advantages of HRMA in routine medical microbiology for identification, strain typing and further characterization of pathogenic bacteria and fungi in particular. The results show increasing numbers of newly-developed and more tailor-made assays in this field. For microbiologists unfamiliar with technical aspects of HRMA, we also provide insight into the technique from the perspective of microbial characterization.Conclusions. We can anticipate that the routine availability of HRMA in medical microbiology laboratories will provide a strong stimulus to this field. This is already envisioned by the growing number of medical microbiology applications published recently. The speed, power, convenience and cost effectiveness of this technology virtually predestine that it will advance genetic characterization of microbes and streamline, facilitate and enrich diagnostics in routine medical microbiology without interfering with the proven advantages of conventional cultivation.

SLCO1B3 screening in colorectal cancer patients using High-Resolution Melting Analysis method and immunohistochemistry

“…To our knowledge, this is the first study of SLCO1B3 gene using HRMA in FFPE tissue from patients with CRC and in comparison with immunohistochemistry. The interpretation of the results was difficult because of the limited number of samples (30). Accurate determination of specificity requires analysis of large numbers of known wild type samples.…”

“…avoid long fragments and multiple melt domains), designing a HRMA screen for a gene is rather straightforward. [28][29][30][31] Moreover, HRMA seems to represent a more sensitive approach to detect a minimal fraction of mutated cells in neoplastic tissue that, in many cases, is not achievable by direct sequencing. 32 Among the many applications of HRMA method, HRMA provides a convenient way to detect SNP mutations in a targeted gene without performing full fragment sequencing.…”

Theoretical and Experimental Investigations of DNA Open States

“…Профили, получаемые методом HRMA, чувствительны даже к единичным заменам пар оснований. Подобный анализ используют для выявления мутаций, генотипирования возбудителей различных заболеваний и в других областях биологии и медицины [123][124][125][126]. Пример дифференциальных профилей плавления, получаемых при помощи HRMA, представлен на рис.…”

Rapid Detection and Identification of Nontuberculous Mycobacterial Pathogens in Fish by Using High-Resolution Melting Analysis

“…High-resolution melting analysis (HRMA), further developed from real-time PCR, is an emerging technique in medical microbiology that may allow simultaneous detection and diagnosis of pathogens at the species and subspecies levels (24)(25)(26). This technique, first reported in 2002, is based on the difference in melting behaviors of DNA molecules, according to their sequence, lengths, and GC content (25,27). HRMA requires only nanogram amounts of DNA and has the potential to discriminate closely related microorganisms with high accuracy, speed, and sensitivity.…”