Hosseini-Safa Ahmad, Shojaee Saeedeh, Salami Seyed Alireza, Mohebali Mehdi, Hantoushzadeh Sedigheh, Mousavi Parisa, Dehghan Manshadi Seyed Ali, Keshavarz Valian Hossein
Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Department Biotechnology, School of Agricultural Sciences and Engineering, University of Tehran, Tehran, Iran.
Iran J Public Health. 2020 Oct;49(10):1983-1991. doi: 10.18502/ijph.v49i10.4704.
is an obligate intracellular protozoan with worldwide distribution. Diagnosis of toxoplasmosis is a very critical issue, especially in pregnant women and immunocompromised patients. The aim of this study was rapid detection of DNA in peripheral blood samples (PBS) employing HRM technique and using RE gene.
Totally, 242 samples from pregnant women and human immunodeficiency virus (HIV) patients were collected from different hospitals and medical centers of Tehran during Oct 2017 to Dec 2018. High resolution melting analysis (HRM) using partial sequences of repetitive element (RE) gene was done and compared with ELISA test.
Overall, 51 were positive for acute toxoplasmosis that among them, 12 and 20 reported as positive in pregnant women and HIV patients, respectively using HRM technique. Among 70 patients in chronic phase of disease, 10 and 3 samples were reported as positive for pregnant women and HIV patients respectively. From 121 negative control, 3 (4.62%) samples associated with HIV patients, showed positive real-time PCR and HRM analysis results.
For the first time, HRM technique via employing RE gene was used for detection of infection in PBS. This method is suitable, helpful and in parallel with serological methods for early diagnosis of acute as well as active form of toxoplasmosis in pregnant women and HIV patients. The use of techniques based on melt curve and through employing next-generation dyes for diagnosis of would be accessible for patients in developing countries.
是一种专性细胞内原生动物，分布于世界各地。弓形虫病的诊断是一个非常关键的问题，尤其是在孕妇和免疫功能低下的患者中。本研究的目的是采用高分辨率熔解曲线分析（HRM）技术并利用重复元件（RE）基因快速检测外周血样本（PBS）中的弓形虫DNA。
2017年10月至2018年12月期间，从德黑兰不同医院和医疗中心收集了242份孕妇和人类免疫缺陷病毒（HIV）患者的样本。使用重复元件（RE）基因的部分序列进行高分辨率熔解分析（HRM），并与酶联免疫吸附测定（ELISA）试验进行比较。
总体而言，51例急性弓形虫病呈阳性，其中使用HRM技术分别有12例孕妇和20例HIV患者被报告为阳性。在70例疾病慢性期患者中，分别有10例孕妇和3例HIV患者被报告为阳性。在121份阴性对照中，3份（占4.62%）与HIV患者相关的样本，实时聚合酶链反应（PCR）和HRM分析结果呈阳性。
首次使用基于RE基因的HRM技术检测PBS中的弓形虫感染。该方法适用于孕妇和HIV患者急性以及活动期弓形虫病的早期诊断，与血清学方法并行，具有帮助作用。基于熔解曲线并通过使用新一代染料的技术用于弓形虫病诊断，发展中国家的患者也能够使用。
