title: Comparison of loop-mediated isothermal amplification (LAMP) and nested-PCR assay targeting the RE and B1 gene for detection of Toxoplasma gondii in blood samples of children with leukaemia.
abstract: Toxoplasmosis diagnosis constitutes an important measure for disease prevention and control. In this paper, a newly described DNA amplification technique, loop-mediated isothermal amplification (LAMP), and nested-PCR targeting the repeated element (RE) and B1 gene, were compared to each other for the detection of Toxoplasma gondii DNA in blood samples of children with leukaemia. One hundred ten blood samples from these patients were analyzed by LAMP and nested-PCR. Out of 50 seropositive samples (IgM+, IgG+), positive results were obtained with 92% and 86% on RE, B1-LAMP and 82% and 68% on RE, B1-nested PCR analyses, respectively. Of the 50 seronegative samples, three, two and one samples were detected positive by RE-LAMP, B1-LAMP and RE-nested PCR assays, respectively, while none were detected positive by B1-nested PCR. None of the 10 IgM-, IgG+ samples was detected positive after testing LAMP and nested-PCR assays in duplicate. This is the first report of a study in which the LAMP method was applied with high sensitivity and efficacy for the diagnosis of T. gonii in blood samples of children with leukaemia.
authors.affiliation: Department of Medical Parasitology and Mycology, Lorestan University of Medical Sciences, Khorramabad, Iran; Department of Medical Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Shirzad
authors.initials: S
authors.lastname: Fallahi
authors.affiliation: Department of Medical Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Seyyed Javad
authors.initials: SJ
authors.lastname: Seyyed Tabaei
authors.affiliation: Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
authors.forename: Yadollah
authors.initials: Y
authors.lastname: Pournia
authors.affiliation: Department of Medical Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Nozhat
authors.initials: N
authors.lastname: Zebardast
authors.affiliation: Department of Medical Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Biotechnology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address: Bahram_14@yahoo.com.
authors.forename: Bahram
authors.initials: B
authors.lastname: Kazemi
mesh_terms: D000818:Animals; D001769:Blood / Q000469:parasitology*; D002648:Child; D002675:Child, Preschool; D016054:DNA, Protozoan / Q000097:blood*; D006801:Humans; D007938:Leukemia / Q000150:complications*; D051379:Mice; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014123:Toxoplasmosis / Q000175:diagnosis*
chemical_list: D016054:DNA, Protozoan
keywords: B1; Children with leukaemia; LAMP; Nested-PCR; RE; Toxoplasma gondii