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Comparison of the Re-529 Sequence and B1 Gene for Toxoplasma Gondii Detection in Blood Samples of the At-Risk Seropositive Cases Using Uracil Dna Glycosylase Supplemented Loop-Mediated Isothermal Amplification (Udg-Lamp) Assay Publisher Pubmed



Valian HK1 ; Mirhendi H2 ; Mohebali M1, 3 ; Shojaee S1 ; Fallahi S4, 5 ; Jafari R6 ; Kheirandish F5 ; Mousavi P7
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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 of Medical Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
  3. 3. Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran
  4. 4. Hepatitis Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran
  5. 5. Department of Medical Parasitology and Mycology, Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran
  6. 6. Department of Medical Parasitology and Mycology, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran
  7. 7. Skin Diseases and Leishmaniasis Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

Source: Microbial Pathogenesis Published:2020


Abstract

Diagnosis of toxoplasmosis is an important issue, especially in at-risk patients. The molecular methods showed a promising future for such diagnosis; however, the method itself and the target sequence to be detected is an important part of accurate detection of the infection. The aim of the present study was to evaluate the RE-529 sequence and B1 gene for Toxoplasma gondii detection in blood samples of the at-risk seropositive cases using uracil DNA glycosylase supplemented loop-mediated isothermal amplification (UDG-LAMP) assay. In this study, 110 T. gondii seropositive at-risk individuals (pregnant women and immunocompromised patients) and 110 seronegative controls were enrolled. The two most studied sequences (RE-529 and B1) were used and compared for accurate and reliable detection of T. gondii in blood samples using UDG-LAMP assay and compared with real-time PCR method. The detection limit, accuracy, and reliability of UDG-LAMP for the parasite's DNA were also studied. Among 110 studied cases, 39 (35.45%) and 36 (32.7%) were positive for T. gondii DNA with the RE-LAMP and B1-LAMP, respectively. The seronegative cases remained negative for T. gondii DNA with the studied genes, however, there were few false negatives compared with real-time PCR method. The detection limit of the UDG-LAMP for both DNA targets was 0.16 tachyzoite's DNA per reaction tube. Based on the results of this study, the RE-529 sequence has a better detection rate compared to the B1 gene for toxoplasmosis among at-risk people. UDG-LAMP is a highly sensitive, accurate, and reliable method with no false-positive results for the diagnosis of T. gondii infection in blood specimens, however few cases may be missed. © 2019
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