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

Fallahi, Shirzad and Arab Mazar, Zahra and Ghasemian, Mehrdad and Haghighi, Ali (2015) Challenging loop-mediated isothermal amplification (LAMP) technique for molecular detection of Toxoplasma gondii. Asian Pacific Journal of Tropical Medicine. pp. 366-372.

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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.

Item Type: Article
Subjects: R Medicine > R Medicine (General)
Depositing User: samira sepahvandy
Date Deposited: 25 Sep 2016 07:15
Last Modified: 25 Sep 2016 07:15
URI: http://eprints.lums.ac.ir/id/eprint/199

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