Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
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DOI：
10.3390/pathogens8030152
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发表时间：
2019-09-01
期刊：
影响因子：
3.7
通讯作者：
Nuchprayoon, Surang
中科院分区：
文献类型：
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作者：
Sanprasert, Vivornpun;Kerdkaew, Ruthairat;Nuchprayoon, Surang
土壤传播蠕虫（STH）是世界范围内感染人类的最常见肠道寄生虫。STH感染是发病和残疾的主要原因。准确的诊断工具对于评估寄生虫感染的确切患病率至关重要。显微镜检查和培养技术已用于观察粪便样本中是否存在寄生虫卵或幼虫，但它们耗时且灵敏度低。因此，准确，简单，廉价的诊断技术仍然需要同时检测STH感染。虽然已经开发了基于分子的技术，如实时PCR和多重实时PCR，但由于需要昂贵的试剂和仪器，它们不适合常规诊断。在这项研究中，我们建立了一个传统的多重PCR同时快速检测蛔虫，美洲钩虫，粪类圆线虫标本。我们的研究结果表明，多重PCR可以在很低的目标基因浓度（低于1 pg）检测STH的DNA，没有交叉扩增。多重PCR检测多重感染的灵敏度是福尔马林-乙酸乙酯浓缩法的5倍，检测沙门氏菌的灵敏度是福尔马林-乙酸乙酯浓缩法的2倍。粪虫但多重PCR检测A.蚓类和N.美国人该方法可以作为一种检测STH的替代方法，特别是对S。粪虫
Soil-transmitted helminths (STHs) are the most common intestinal parasites infecting humans worldwide. STH infections are a major cause of morbidity and disability. Accurate diagnostic tools are pivotal for assessing the exact prevalence of parasitic infections. Microscopic examination and culture techniques have been used to observe the presence of eggs or larvae of parasites in stool samples, but they are time-consuming and have low sensitivity. Therefore, accurate, simple, and inexpensive diagnostic techniques are still required for simultaneous detection of STH infections. Although molecular-based techniques, such as real-time PCR and multiplex real-time PCR, have been developed, they are not suitable for routine diagnosis due to the requirement for expensive reagents and instruments. In this study, we established a conventional multiplex PCR for simultaneous rapid detection of Ascaris lumbricoides, Necator americanus, and Strongyloides stercoralis in stool samples. Our results show that the multiplex PCR could detect the DNA of STHs at a very low target gene concentrations (lower than 1 pg) with no cross-amplification. Multiplex PCR had five times higher sensitivity than the formalin-ethyl acetate concentration technique (FECT) in the detection of multiple infections, and two times higher for detection of S. stercoralis. However, multiplex PCR was comparable to FECT in the detection of A. lumbricoides and N. americanus. In conclusion, this method could be used as an alternative method for the detection of STHs, especially for S. stercoralis.