Challenging loop-mediated isothermal amplification(LAMP) technique for molecular detection of Toxoplasma gondii

作     者：Shirzad Fallahi Zahra Arab Mazar Mehrdad Ghasemian Ali Haghighi 

作者机构：Department of Medical Parasitology and MycologySchool of MedicineLorestan University of Medical Sciences Department of Medical Parasitology and MycologySchool of MedicineShahid Beheshti University of Medical Sciences Department of Medical Parasitology and MycologySchool of MedicineIran University of Medical Sciences 

出 版 物：《Asian Pacific Journal of Tropical Medicine》 (亚太热带医药杂志（英文版）)

年 卷 期：2015年第8卷第5期

页      面：366-372页

核心收录：

学科分类：100208[医学-临床检验诊断学] 1002[医学-临床医学] 10[医学] 

基　　金：supported financially by grant of Lorestan University of Medical Sciences Khorramabad Iran 

主　　题：LAMP Nested-PCR Molecular detection Toxoplasma gondii 

摘      要：Objective:To compare analytical sensitivity and specificity of a newly described DNA amplification technique.LAMP and nested PCR assay targeting the RE and Bl genes for the detection of Toxoplasma gondii(T.gondii) DNA.Methods:The analytical sensitivity of LAMP and ncstcd-PCR was obtained against 10-fold serial dilutions of T.gondii DNA ranging from 1 ng to 0.01 fg.DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays.Results:After testing LAMP and nesled-PCR in duplicate,the detection limit of RE-LAMP.B1-LAMP,RE-nested PCR and B1-nested PCR assays was one fg.100 fg,1 pg and 10 pg of T.gondii DNA respectively.All the LAMP assays and nested PCRs were 100% specific.The RE-LAMP assay revealed the most sensitivity for the detection of T.gondii DNA.Conclusions:The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T.gondii.Furthermore,these findings indicate that primers based on the RE are more suitable than those based on the B1 gene.However,the B1-LAMP assay has potential as a diagnostic tool for detection of T.gondii.