Abstract:
:Several DNA probes were designed for use in real-time polymerase chain reaction (PCR) assays to target sequence variation within the ribosomal intergenic spacer (IGS) of schistosomes. A sub-section of the IGS (∼300bp) was amplified, with cross-specific primers, after which group-specific fluorescent, locked nucleic acid probes were assessed for their ability to differentiate and quantify DNA from Schistosoma haematobium and Schistosoma mansoni group parasites. A number of fluorescent probe candidates were screened and validated against genomic DNA from adult schistosome worms and laboratory infected freshwater snails. Two fluorescent, locked nucleic acid probes ShaemLNA5 and SmanLNA2, of 20-26bp in length, were identified and found to be effective in providing evidence of infection in field-collected snails. To adapt these real-time PCR assays for more resource-poor laboratory settings, a PCR-restriction fragment length polymorphism (RFLP) assay was developed and primer/probe combinations were modified for use in oligochromatography, a DNA 'dipstick' technology. An appropriate dipstick was developed, inclusive of internal amplification and amplicon migration controls that could be of particular importance for assessing schistosome transmission dynamics. These assays and tools also have future potential for use in detection of schistosome infections in humans and livestock.
journal_name
Acta Tropjournal_title
Acta tropicaauthors
Kane RA,Stothard JR,Rollinson D,Leclipteux T,Evraerts J,Standley CJ,Allan F,Betson M,Kaba R,Mertens P,Laurent Tdoi
10.1016/j.actatropica.2011.10.019subject
Has Abstractpub_date
2013-11-01 00:00:00pages
241-9issue
2eissn
0001-706Xissn
1873-6254pii
S0001-706X(11)00315-9journal_volume
128pub_type
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