El-Kareem Nasser Mohamed Abd, Dyab Ahmed Kamal, Albalawi Nada Oudah, El Samea Abdalla Abd, Taha Mohamed Ahmed Ali, AlQadeeb Hajar, Gareh Ahmed, Hiekal Elham Adel, Alzaylaee Hind, Elmahallawy Ehab Kotb
Department of Medical Parasitology, Faculty of Medicine, Al-Azhar University (Assiut Branch), Assiut, Egypt.
Department of Medical Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt.
Front Microbiol. 2024 Nov 20;15:1499270. doi: 10.3389/fmicb.2024.1499270. eCollection 2024.
Trichomoniasis remains one of the most significant sexually transmitted disease (STDs) for public health. The disease is caused by parasitic protozoa, (), which is often underestimated in tropical medicine. Despite its public health importance, the epidemiology and molecular characteristics of trichomoniasis in Egypt remains poorly understood, particularly in the southern part of the country (Upper Egypt). This study targeted exploring the genetic variability of infections in Egyptian women living in Upper Egypt using restriction fragment length polymorphism (RFLP).
This cross-sectional study included 150 female patients, who visited the gynaecology and obstetrics outpatient clinics at Sohag General Hospital between 2019 and 2022, exhibiting symptoms of trichomoniasis. Vaginal washout samples were collected from each patient and analyzed using three diagnostic techniques: direct wet mount microscopy, culture on TYM Diamond's medium, and PCR amplification and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) targeting the actin gene, which was applied to all 16 samples that tested positive in culture. The PCR-RFLP results were then visualized through agarose gels electrophoresis to detect DNA fragments.
Out of 150 vaginal washout samples, 12 cases (8%) tested positive for trophozoites via direct wet mount microscopy, while 16 samples (10.6%) were positive in culture. Additionally, PCR-RFLP analysis of the 16 culture-positive samples revealed that 13 samples were confirmed positive using this molecular method. The amplified products were digested with the restriction enzyme Hind II, yielding three DNA fragments of 60, 213, and 827 bp, which were then detected by agarose gel electrophoresis. Digestion with RsaI produced five fragments measuring 87, 103/106, 236, and 568 bp, while MseI digestion resulted in three distinct fragments of 204, 315, and 581 bp.
This study provides robust baseline data on the prevalence and microscopic characteristics of in Upper Egypt, while also presenting, for the first time, molecular detection and genotyping and revealed that genotype E is the only prevalent genotype in the region.
滴虫病仍然是对公共卫生具有重要意义的性传播疾病之一。该疾病由寄生原生动物()引起,在热带医学中常常被低估。尽管其对公共卫生很重要,但埃及滴虫病的流行病学和分子特征仍知之甚少,尤其是在该国南部地区(上埃及)。本研究旨在利用限制性片段长度多态性(RFLP)探索生活在上埃及的埃及女性中感染的基因变异性。
这项横断面研究纳入了150名女性患者,她们在2019年至2022年期间前往索哈杰综合医院的妇产科门诊就诊,表现出滴虫病症状。从每位患者收集阴道冲洗样本,并使用三种诊断技术进行分析:直接湿片显微镜检查、在TYM戴蒙德培养基上培养,以及针对肌动蛋白基因的聚合酶链反应扩增和聚合酶链反应 - 限制性片段长度多态性(PCR - RFLP),该方法应用于所有在培养中检测呈阳性的16个样本。然后通过琼脂糖凝胶电泳观察PCR - RFLP结果以检测DNA片段。
在150个阴道冲洗样本中,12例(8%)通过直接湿片显微镜检查检测到滋养体呈阳性,而16个样本(10.6%)培养呈阳性。此外,对16个培养阳性样本的PCR - RFLP分析显示,使用这种分子方法确认其中13个样本呈阳性。扩增产物用限制性内切酶Hind II消化,产生60、213和827 bp的三个DNA片段,然后通过琼脂糖凝胶电泳检测到。用RsaI消化产生五个片段,大小分别为87、103/106、236和568 bp,而用MseI消化产生204、315和581 bp的三个不同片段。
本研究提供了关于上埃及地区滴虫病患病率和微观特征的有力基线数据,同时首次展示了分子检测和基因分型,并揭示基因型E是该地区唯一流行的基因型。