Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and melting curve analysis
Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and melting curve analysis
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基于实时荧光共振能量转移PCR的方法和熔解曲线分析检测受感染的螺类中的后睾虫
DOI：
10.1007/s00436-008-1026-0
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发表时间：
2008
影响因子：
2
通讯作者：
W. Maleewong
中科院分区：
文献类型：
--
作者：
P. Intapan;T. Thanchomnang;V. Lulitanond;P. Pongsaskulchoti;W. Maleewong
建立了一种实时荧光共振能量转移（FRET）PCR结合熔解曲线分析检测实验感染的第一中间宿主--灵猫后睾吸虫的方法。该试验基于来自pOV-A6特异性探针序列的扩增子与O.灵猫和特异性荧光团标记的探针。实时FRET PCR可以检测到一个单一的尾蚴人工引入池的30个未感染的蜗牛。手术通过它们的解链温度，可以将被灵猫感染的蜗牛与未被感染的蜗牛以及其他寄生虫DNA的基因组DNA区分开。该方法的敏感性和特异性均为100%。解链曲线分析法是一种灵敏的O。它是快速的，允许高通量，并且可以在小样品上完成。该方法不仅对O.在监测传播场所时，还可用于检测自然水道中的尾蚴感染。
A real-time fluorescence resonance energy transfer (FRET) PCR combined with melting curve analysis was developed for the detection of Opisthorchis viverrini in experimentally infected bithynid snails, its first intermediate hosts. The test is based on the fluorescence melting curve analysis of a hybrid between an amplicon from the pOV-A6-specific probe sequence, a 162-bp repeated sequence specific to O. viverrini and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single cercaria artificially introduced in a pool of 30 non-infected snails. The O. viverrini-infected snails were discriminated from non-infected snails and from genomic DNA of other parasite DNAs by their melting temperatures. Sensitivity and specificity of this method were both 100%. Melting curve analysis is a sensitive alternative for the specific detection of O. viverrini-infected snails; it is rapid, allows a high throughput, and can be done on small samples. The assay not only has a high potential for epidemiological surveys of O. viverrini-infected bithynid snails, but also for the detection of cercariae infestations of natural waterways when monitoring transmission sites.
DOI：
10.4269/ajtmh.2007.76.950
发表时间：
2007-05-01
影响因子：
3.3
作者：
Abbasi, Ibrahim;King, Charles H.;Hamburger, Joseph
通讯作者：
Hamburger, Joseph