Rapid detection of Opisthorchis viverrini copro-DNA using loop-mediated isothermal amplification (LAMP).
Rapid detection of Opisthorchis viverrini copro-DNA using loop-mediated isothermal amplification (LAMP).
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DOI：
10.1016/j.parint.2011.08.009
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发表时间：
2012-03
影响因子：
1.9
通讯作者：
Sripa B
中科院分区：
文献类型：
--
作者：
Arimatsu Y;Kaewkes S;Laha T;Hong SJ;Sripa B
猫尾后睾吸虫和其他食源性吸虫感染是泰国、老挝人民民主共和国、越南和柬埔寨的主要卫生问题。O.在华支睾吸虫和微小肠吸虫共存的地区，很难根据寄生虫卵的显微镜观察来鉴定灵猫吸虫。建立了一种快速、灵敏、特异的O.使用环介导等温扩增（LAMP）方法从粪便样品中检测灵猫病毒感染。从7个区域共设计了5条引物，以核糖体DNA的内转录间隔区1（ITS 1）为靶点进行特异性扩增。与实时荧光法和浊度法相比，羟基萘酚蓝（HNB）检测方法简单、特异性强，是一种更有效的检测LAMP产物的方法。LAMP检测特异性扩增O. viverrini ITS 1但不是C.中华绒螯蟹和微小肠吸虫，检测限约为10−3 ng DNA/μL。与鸡卵阳性样品相比，LAMP的灵敏度为100%。虽然所有显微镜下阳性样本均为LAMP阳性，但另外5/13份（38.5%）显微镜下阴性样本也为LAMP阳性。该技术在混合型O.灵猫和肠道吸虫感染。由于这是一种容易和简单的方法，LAMP可能适用于床旁诊断。
Opisthorchis viverrini and other foodborne trematode infections are major health problem in Thailand, the Lao People’s Democratic Republic, Vietnam and Cambodia. Differential diagnosis of O. viverrini based on the microscopic observation of parasite eggs is difficult in areas where Clonorchis sinensis and minute intestinal flukes coexist. We therefore established a rapid, sensitive and specific method for detecting O. viverrini infection from the stool samples using the loop-mediated isothermal amplification (LAMP) method. A total of five primers from seven regions were designed to target the internal transcribed spacer 1 (ITS1) in ribosomal DNA for specific amplification. Hydroxy naphthol blue (HNB) was more effective to detect the LAMP product compared to the Real-time LAMP and turbidity assay for its simple and distinct detection. The LAMP assay specifically amplified O. viverrini ITS1 but not C. sinensis and minute intestinal flukes with the limit of detection around 10−3 ng DNA/μL. The sensitivity of the LAMP was 100% compare to egg positive samples. While all microscopically positive samples were positive by LAMP, additionally 5 of 13 (38.5%) microscopically negative samples were also LAMP positive. The technique has great potential for differential diagnosis in endemic areas with mixed O. viverrini and intestinal fluke infections. As is is an easy and simple method, the LAMP is potentially applicable for point-of-care diagnosis.
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影响因子：
15.8
作者：
Sripa, Banchob;Kaewkes, Sasithorn;Sithithaworn, Paiboon;Mairiang, Eimorn;Laha, Thewarach;Smout, Michael;Pairojkul, Chawalit;Bhudhisawasdi, Vajaraphongsa;Tesana, Smarn;Thinkamrop, Bandit;Bethony, Jeffrey M.;Loukas, Alex;Brindley, Paul J.
通讯作者：
Brindley, Paul J.
影响因子：
2.4
作者：
Maleewong, W;Intapan, PM;Kitikoon, V
通讯作者：
Kitikoon, V
一种新的基于PCR的方法表明Clonorchis sinensis的范围现在延伸到泰国中部。
影响因子：
3.8
作者：
Traub RJ;Macaranas J;Mungthin M;Leelayoova S;Cribb T;Murrell KD;Thompson RC
通讯作者：
Thompson RC
影响因子：
--
作者：
Maeda, H;Kokeguchi, S;Takashiba, S
通讯作者：
Takashiba, S
需要协调一致的动作来解决亚洲的肝氟化物感染。
影响因子：
3.8
作者：
Sripa B
通讯作者：
Sripa B