生物
量油尺
放大器
聚合酶链反应
核糖体RNA
分子生物学
核糖体DNA
实时聚合酶链反应
限制性片段长度多态性
基因组DNA
基因间区
塔克曼
DNA
遗传学
基因
基因组
生物化学
系统发育学
尿
作者
Richard A. Kane,J. Russell Stothard,David Rollinson,T. Leclipteux,Jonathan Evraerts,Claire J. Standley,Fiona Allan,Martha Betson,Rehana Kaba,Pascal Mertens,Thierry Laurent
出处
期刊:Acta Tropica
[Elsevier]
日期:2013-11-01
卷期号:128 (2): 241-249
被引量:33
标识
DOI:10.1016/j.actatropica.2011.10.019
摘要
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 (∼300 bp) 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 20–26 bp 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.
科研通智能强力驱动
Strongly Powered by AbleSci AI