A high resolution melting (HRM) technology-based assay for cost-efficient clinical detection and genotyping of herpes simplex virus (HSV)-1 and HSV-2

Abstract

Genital herpes can be caused by two very similar viruses, herpes simplex virus (HSV)-1 or HSV-2. These two HSV types cannot be distinguished clinically, but genotyping is recommended in the first-episodes of genital herpes to guide counselling and management. Quantitative polymerase chain reaction (qPCR) is the preferred diagnostic method for HSV typing. However, commercial qPCR methods use expensive fluorescent labeled probes for detection. Furthermore, most low-cost methods are not able to differentiate between HSV-1 and -2. The aim of this study was to develop a high resolution melting (HRM) technology-based assay for sensitive HSV-1 and HSV-2 detection and genotyping. Using a panel of 46 clinical specimens, the performance of the HRM assay was compared to two commercial HSV tests: the HRM assay detected HSV in all 23 positive samples, with no false positive results (100% concordance with HSV I/II Real-TM assay). Additionally, the HRM assay correctly genotyped both HSV types in a subset of these clinical samples, as determined by the Realstar HSV PCR Kit. The HSV HRM assay provides a cost-effective alternative method to conventional more expensive assays and can be used in routine clinical specimens, in cases where it is particularly necessary to detect and distinguish HSV-1 from -2.

Detection and Differentiation of Two Koala Gammaherpesviruses by Use of High-Resolution Melt (HRM) Analysis Reveals Differences in Viral Prevalence and Clinical Associations in a Large Study of Free-Ranging Koalas

“…We used high-resolution melt analysis of amplicons generated by a widely used pan-herpesvirus nested PCR to differentiate between the two koala gammaherpesviruses, PhaHV-1 and PhaHV-2. HRM analysis has been used previously to differentiate between related strains of other veterinary and human pathogens (27)(28)(29)(30). As this assay targets a conserved region of the herpesvirus DNA polymerase gene, similar approaches could also be suitable for other marsupial herpesviruses in other host species.…”

Herpes Simplex Virus Type-2 Paralyzes the Function of Monocyte-Derived Dendritic Cells

“…Primary HSV-1 infection generally occurs during early childhood and is usually an asymptomatic inapparent orolabial infection, via direct contact of infectious lesions/body fluids [66,67]. HSV-2 infections are often associated with transient genital lesions, since HSV-2 is among the most frequent sexually transmitted infections, and often linked with a higher risk for HIV acquisition [68,69]. However, during the last years it has become clear that the classification between HSV-1-caused orofacial herpes versus HSV-2-caused genital herpes is not as strict as supposed, since both viruses are able to induce similar symptoms [70,71].…”

The development of unlabeled probes-high resolution melting (UP-HRM) marker on SAD, IAA27 and ACC genes of oil palm

“…The advantage of UP-HRM method is not required fluorescent labels on their special primers or enzymes. UP-HRM is most costeffective compared to other genotyping technologies such as single-pass sequencing genotyping (Zhang et al 2013;Lieveld et al 2017) . UP-HRM is fast and strong, allowing rapid genotyping of many samples.…”