Department of Tropical Medicine, Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul, 03722, Republic of Korea.
Sci Rep. 2024 Oct 23;14(1):25049. doi: 10.1038/s41598-024-76304-1.
Recent advancements in next-generation sequencing (NGS) technologies have created new opportunities for comprehensive screening of multiple parasite species. In this study, we cloned the 18 S rDNA V9 region of 11 species of intestinal parasites into plasmids. Equal amounts and concentrations of these 11 plasmids were pooled, and amplicon NGS targeting the 18 S rDNA V9 region was performed using the Illumina iSeq 100 platform. A total of 434,849 reads were identified, and all 11 parasite species were detected, although the number of output reads for each parasite varied. The read count ratio, in descending order, was as follows: Clonorchis sinensis, 17.2%; Entamoeba histolytica, 16.7%; Dibothriocephalus latus, 14.4%; Trichuris trichiura, 10.8%; Fasciola hepatica, 8.7%; Necator americanus, 8.5%; Paragonimus westermani, 8.5%; Taenia saginata, 7.1%; Giardia intestinalis, 5.0%; Ascaris lumbricoides, 1.7%; and Enterobius vermicularis, 0.9%. We found that the DNA secondary structures showed a negative association with the number of output reads. Additionally, variations in the amplicon PCR annealing temperature affected the relative abundance of output reads for each parasite. These findings can be applied to improve parasite detection methodologies and ultimately enhance efforts to control and prevent intestinal parasitic infections.
近年来，下一代测序（NGS）技术的进步为全面筛选多种寄生虫物种创造了新的机会。在这项研究中，我们将 11 种肠道寄生虫的 18S rDNA V9 区克隆到质粒中。将这些 11 个质粒等量且浓度混合，然后使用 Illumina iSeq 100 平台对 18S rDNA V9 区进行扩增子 NGS。共鉴定出 434849 个读数，所有 11 种寄生虫均被检测到，尽管每种寄生虫的输出读数数量不同。按降序排列，读计数比如下：华支睾吸虫，17.2%；溶组织内阿米巴，16.7%；阔节裂头绦虫，14.4%；毛首鞭形线虫，10.8%；肝片形吸虫，8.7%；美洲板口线虫，8.5%；卫氏并殖吸虫，8.5%；猪带绦虫，7.1%；肠贾第鞭毛虫，5.0%；蛔虫，1.7%；和蛲虫，0.9%。我们发现 DNA 二级结构与输出读数呈负相关。此外，扩增子 PCR 退火温度的变化会影响每种寄生虫的输出读数的相对丰度。这些发现可用于改进寄生虫检测方法，并最终加强控制和预防肠道寄生虫感染的努力。
