Molecular Identification of Echinococcus spp. and Other Taeniid Tapeworms Using Next-Generation Sequence Analysis of PCR Amplified 18s rRNA Gene  

作　　者：Rasmi Abu-Helu George Kokaly Sajeda Nojoum Imad Matouk Murad Ibrahim Ibrahim Abbasi Rasmi Abu-Helu;George Kokaly;Sajeda Nojoum;Imad Matouk;Murad Ibrahim;Ibrahim Abbasi(Department of Medical Laboratory Sciences, Faculty of Health Professions, Al-Quds University, East Jerusalem;Department of Microbiology and Immunology, Faculty of Medicine, Al-Quds University, East Jerusalem;Department of Biological Sciences, Al-Quds University, East Jerusalem)

机构地区：[1]Department of Medical Laboratory Sciences, Faculty of Health Professions, Al-Quds University, East Jerusalem [2]Department of Microbiology and Immunology, Faculty of Medicine, Al-Quds University, East Jerusalem [3]Department of Biological Sciences, Al-Quds University, East Jerusalem

出　　处：《American Journal of Molecular Biology》2025年第1期75-87,共13页美国分子生物学期刊(英文)

摘　　要：Cystic echinococcosis (CE) is a prevalent zoonotic disease caused by Echinococcus granulosus, with a cosmopolitan distribution. The parasite is transmitted cyclically between canines and numerous intermediate herbivorous livestock animals. Also, other Taeniid tapeworms could infect domestic dogs and they pose significant veterinary and public health concerns worldwide. This study aimed to develop a sensitive molecular method for detecting Echinococcus spp. DNA in dog fecal samples using next-generation sequencing (NGS). A set of PCR primers targeting conserved regions of Taeniid tapeworms’ 18s rRNA genes was designed and tested for amplifying genomic DNA from various tapeworm species. The PCR system demonstrated high sensitivity, amplifying DNA from all tested tapeworm species, with differences observed in amplified band sizes. The primers were adapted for NGS analysis by adding forward and reverse adapters, enabling the sequencing of amplified DNA fragments. Application of the developed PCR system to dog fecal samples collected from Yatta town, Palestine, revealed the presence of E. granulosus DNA in five out of 50 samples. NGS analysis confirmed the specificity of the amplified DNA fragments, showing 98% - 99% similarity with the 18s rDNA gene of E. granulosus. This study demonstrates the utility of NGS-based molecular methods for accurate and sensitive detection of Echinococcus spp. in dog fecal samples, providing valuable insights for epidemiological surveillance and control programs of echinococcosis in endemic regions.Cystic echinococcosis (CE) is a prevalent zoonotic disease caused by Echinococcus granulosus, with a cosmopolitan distribution. The parasite is transmitted cyclically between canines and numerous intermediate herbivorous livestock animals. Also, other Taeniid tapeworms could infect domestic dogs and they pose significant veterinary and public health concerns worldwide. This study aimed to develop a sensitive molecular method for detecting Echinococcus spp. DNA in dog fecal samples using next-generation sequencing (NGS). A set of PCR primers targeting conserved regions of Taeniid tapeworms’ 18s rRNA genes was designed and tested for amplifying genomic DNA from various tapeworm species. The PCR system demonstrated high sensitivity, amplifying DNA from all tested tapeworm species, with differences observed in amplified band sizes. The primers were adapted for NGS analysis by adding forward and reverse adapters, enabling the sequencing of amplified DNA fragments. Application of the developed PCR system to dog fecal samples collected from Yatta town, Palestine, revealed the presence of E. granulosus DNA in five out of 50 samples. NGS analysis confirmed the specificity of the amplified DNA fragments, showing 98% - 99% similarity with the 18s rDNA gene of E. granulosus. This study demonstrates the utility of NGS-based molecular methods for accurate and sensitive detection of Echinococcus spp. in dog fecal samples, providing valuable insights for epidemiological surveillance and control programs of echinococcosis in endemic regions.

关 键 词：Cystic Echinococcosis Taeniid Tapeworms Next-Generation Sequencing Molecular Detection Dog Fecal Samples 

分 类 号：R73[医药卫生—肿瘤]