Developing a real-time PCR assay based on multiplex high-resolution melt-curve analysis: a pilot study in detection and discrimination of soil-transmitted helminth and schistosome species

Parasitology
Lucas J CunninghamEmily R Adams

Abstract

With the push towards control and elimination of soil-transmitted helminthiasis and schistosomiasis in low- and middle-income countries, there is a need to develop alternative diagnostic assays that complement the current in-country resources, preferably at a lower cost. Here, we describe a novel high-resolution melt (HRM) curve assay with six PCR primer pairs, designed to sub-regions of the nuclear ribosomal locus. Used within a single reaction and dye detection channel, they are able to discriminate Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Ascaris lumbricoides, Trichuris trichiuria and Schistosoma spp. by HRM curve analysis. Here we describe the primers and the results of a pilot assessment whereby the HRM assay was tested against a selection of archived fecal samples from Ghanaian children as characterized by Kato-Katz and real-time PCR analysis with species-specific TaqMan hydrolysis probes. The resulting sensitivity and specificity of the HRM was 80 and 98.6% respectively. We judge the assay to be appropriate in modestly equipped and resourced laboratories. This method provides a potentially cheaper alternative to the TaqMan method for laboratories in lower resource settings. However, the assay...Continue Reading

References

Sep 1, 1994·Molecular & General Genetics : MGG·G J Hunt, R E Page
Apr 12, 2001·Tropical Medicine & International Health : TM & IH·A KongsP Van der Stuyft
Jan 28, 2009·Transactions of the Royal Society of Tropical Medicine and Hygiene·J Russell Stothard
Nov 24, 2012·Journal of Clinical Microbiology·Jie LiuEric R Houpt
Sep 4, 2015·Lancet·Lorenzo SavioliShaali Makame Ame
Jul 25, 2017·Pediatric Clinics of North America·Jill E WeatherheadRojelio Mejia
Nov 9, 2017·Parasitology·Felicia KeesingBrian F Allan

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Citations

May 30, 2020·Nature Reviews. Disease Primers·Kathryn J ElsePhilip J Cooper
Jul 31, 2020·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·Dinesh BalachandraRahmah Noordin
Jun 11, 2020·Tropical Medicine and Infectious Disease·Mirabeau Mbong NgweseAyola Akim Adégnika

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