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Development of High Resolution Melting Analysis As a Diagnostic Tool for Molecular Detection of Toxoplasma Infection in Pregnant Women and Hiv Positive Cases



Hosseinisafa A1 ; Shojaee S1 ; Salami SA2 ; Mohebali M1, 3 ; Hantoushzadeh S4 ; Mousavi P1 ; Dehghan Manshadi SA5 ; Keshavarz Valian H1, 3
Authors
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Authors Affiliations
  1. 1. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Department Biotechnology, School of Agricultural Sciences and Engineering, University of Tehran, Tehran, Iran
  3. 3. Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran
  4. 4. Maternal, Fetal and Neonatal Research Center, Vali-Asr Hospital, Imam Khomeini Hospital Complexes, Tehran University of Medical Sciences, Tehran, Iran
  5. 5. Department of Infectious Diseases and Tropical Medicine and Iranian Research Center for HIV/AIDS, Tehran University of Medical Sciences, Tehran, Iran

Source: Iranian Journal of Public Health Published:2020

Abstract

Background: Toxoplasma gondii 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 T. gondii DNA in peripheral blood samples (PBS) employing HRM technique and using RE gene. Methods: 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. Results: 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. Conclusion: For the first time, HRM technique via employing RE gene was used for detection of T. gondii 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 T. gondii would be accessible for patients in developing countries. © 2020, Iranian Journal of Public Health. All rights reserved.
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