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

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 again...

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Detalles Bibliográficos
Autores principales: Fallahi, Shirzad, Mazar, Zahra Arab, Ghasemian, Mehrdad, Haghighi, Ali
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hainan Medical College. Published by Elsevier (Singapore) Pte Ltd. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7128654/
https://www.ncbi.nlm.nih.gov/pubmed/26003595
http://dx.doi.org/10.1016/S1995-7645(14)60345-X
Descripción
Sumario: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.