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

Shirzad Fallahi; Zahra Arab Mazar; Mehrdad Ghasemian; Ali Haghighi

doi:10.1016/S1995-7645(14)60345-X

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

Asian Pac J Trop Med. 2015 May;8(5):366-72. doi: 10.1016/S1995-7645(14)60345-X.

Authors

Shirzad Fallahi¹, Zahra Arab Mazar², Mehrdad Ghasemian³, Ali Haghighi²

Affiliations

¹ Department of Medical Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran. Electronic address: Shfupdate@gmail.com.
² Department of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
³ Department of Medical Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.

Abstract

Objective: To compare analytical sensitivity and specificity of a newly described DNA amplification technique, LAMP and nested PCR assay targeting the RE and B1 genes for the detection of Toxoplasma gondii (T. gondii) DNA.

Methods: The analytical sensitivity of LAMP and nested-PCR was obtained against10-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 nested-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.

Keywords: LAMP; Molecular detection; Nested-PCR; Toxoplasma gondii.