E. Kaneva, N. Tsvetkova, Desislava Velcheva, R. Harizanov, lskren Kaftandjiev, R. Borisova, M. Videnova, Aleksandra lvanova, I. Alexiev, Reneta Dimitrova, Maria Pavlova
{"title":"开发和应用一种在肠溶病患者样本中对蚯蚓肠杆菌进行基因检测的方法","authors":"E. Kaneva, N. Tsvetkova, Desislava Velcheva, R. Harizanov, lskren Kaftandjiev, R. Borisova, M. Videnova, Aleksandra lvanova, I. Alexiev, Reneta Dimitrova, Maria Pavlova","doi":"10.5272/jimab.2024301.5295","DOIUrl":null,"url":null,"abstract":"Purpose. The routine diagnosis of enterobiasis consists of a microscopic examination of perianal imprints on a transparent scotch tape for the presence of parasitic eggs. However, this method has low sensitivity when the number of eggs is small. The development of highly sensitive and specific methods, such as PCR, will increase the possibilities of detecting the parasite and will enable its genetic characterization. Material/Methods. The study was conducted from May 2022 to September 2022 and comprised 24 patients infected with Enterobius vermicularis. DNA was extracted from parasite eggs/adult worms using PureLink Genomic DNA Mini Kit (INVITROGEN), according to the manufacturer's instructions. Nested polymerase chain reaction (PCR) for detection of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was applied with two sets of primers. Results. The developed and described method for genetic determination of Enterobius vermicularis in samples from infected individuals is suitable and applicable to improve the diagnosis of the disease, as well as for further phylogenetic research of the parasite, which is of great importance for its successful treatment and control. Conclusions. Regardless of existing morphological diagnostic methods, genotyping studies concerning Enterobius vermicularis need to be deepened. Molecular-biological analyses increase diagnostic sensitivity, provide valuable knowledge about the geographic distribution and diversity of helminths, and allow the monitoring of therapy.","PeriodicalId":508168,"journal":{"name":"Journal of IMAB - Annual Proceeding (Scientific Papers)","volume":"29 9","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"DEVELOPMENT AND APPLICATION OF A METHOD FOR GENETIC DETECTION OF ENTEROBIUS VERMICULARIS IN SAMPLES OF PATIENTS WITH ENTEROBIАSIS\",\"authors\":\"E. Kaneva, N. Tsvetkova, Desislava Velcheva, R. Harizanov, lskren Kaftandjiev, R. Borisova, M. Videnova, Aleksandra lvanova, I. Alexiev, Reneta Dimitrova, Maria Pavlova\",\"doi\":\"10.5272/jimab.2024301.5295\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Purpose. The routine diagnosis of enterobiasis consists of a microscopic examination of perianal imprints on a transparent scotch tape for the presence of parasitic eggs. However, this method has low sensitivity when the number of eggs is small. The development of highly sensitive and specific methods, such as PCR, will increase the possibilities of detecting the parasite and will enable its genetic characterization. Material/Methods. The study was conducted from May 2022 to September 2022 and comprised 24 patients infected with Enterobius vermicularis. DNA was extracted from parasite eggs/adult worms using PureLink Genomic DNA Mini Kit (INVITROGEN), according to the manufacturer's instructions. Nested polymerase chain reaction (PCR) for detection of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was applied with two sets of primers. Results. The developed and described method for genetic determination of Enterobius vermicularis in samples from infected individuals is suitable and applicable to improve the diagnosis of the disease, as well as for further phylogenetic research of the parasite, which is of great importance for its successful treatment and control. Conclusions. Regardless of existing morphological diagnostic methods, genotyping studies concerning Enterobius vermicularis need to be deepened. Molecular-biological analyses increase diagnostic sensitivity, provide valuable knowledge about the geographic distribution and diversity of helminths, and allow the monitoring of therapy.\",\"PeriodicalId\":508168,\"journal\":{\"name\":\"Journal of IMAB - Annual Proceeding (Scientific Papers)\",\"volume\":\"29 9\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-01-09\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of IMAB - Annual Proceeding (Scientific Papers)\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.5272/jimab.2024301.5295\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of IMAB - Annual Proceeding (Scientific Papers)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.5272/jimab.2024301.5295","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
目的。肠虫病的常规诊断方法是用显微镜检查透明胶带上的肛周印迹,看是否有寄生虫卵。然而,当虫卵数量较少时,这种方法的灵敏度较低。高灵敏度和特异性方法(如 PCR)的开发将增加检测寄生虫的可能性,并使寄生虫的基因特征得以确定。材料/方法。该研究于 2022 年 5 月至 2022 年 9 月进行,包括 24 名感染蠕虫肠杆菌的患者。根据制造商的说明,使用 PureLink Genomic DNA Mini Kit(INVITROGEN)从寄生虫卵/成虫中提取 DNA。使用两组引物进行巢式聚合酶链反应(PCR)检测线粒体细胞色素 c 氧化酶亚基 1(cox1)基因。结果。所开发和描述的用于确定感染者样本中蠕形肠虫基因的方法适用于改进该疾病的诊断,也适用于寄生虫的进一步系统发育研究,这对成功治疗和控制该疾病具有重要意义。结论无论现有的形态学诊断方法如何,都需要深化有关蠕虫肠杆菌的基因分型研究。分子生物学分析可提高诊断灵敏度,提供有关蠕虫地理分布和多样性的宝贵知识,并可对治疗进行监测。
DEVELOPMENT AND APPLICATION OF A METHOD FOR GENETIC DETECTION OF ENTEROBIUS VERMICULARIS IN SAMPLES OF PATIENTS WITH ENTEROBIАSIS
Purpose. The routine diagnosis of enterobiasis consists of a microscopic examination of perianal imprints on a transparent scotch tape for the presence of parasitic eggs. However, this method has low sensitivity when the number of eggs is small. The development of highly sensitive and specific methods, such as PCR, will increase the possibilities of detecting the parasite and will enable its genetic characterization. Material/Methods. The study was conducted from May 2022 to September 2022 and comprised 24 patients infected with Enterobius vermicularis. DNA was extracted from parasite eggs/adult worms using PureLink Genomic DNA Mini Kit (INVITROGEN), according to the manufacturer's instructions. Nested polymerase chain reaction (PCR) for detection of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was applied with two sets of primers. Results. The developed and described method for genetic determination of Enterobius vermicularis in samples from infected individuals is suitable and applicable to improve the diagnosis of the disease, as well as for further phylogenetic research of the parasite, which is of great importance for its successful treatment and control. Conclusions. Regardless of existing morphological diagnostic methods, genotyping studies concerning Enterobius vermicularis need to be deepened. Molecular-biological analyses increase diagnostic sensitivity, provide valuable knowledge about the geographic distribution and diversity of helminths, and allow the monitoring of therapy.