Challenging loop-mediated isothermal amplification (LAMP) technique for molecular detection ofToxoplasma gondii
Asian Pacific Journal of Tropical Biomedicine
; (12): 366-372, 2015.
Article
en Zh
| WPRIM
| ID: wpr-672826
Biblioteca responsable:
WPRO
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 ofToxoplasma gondii (T. gondii)DNA.Methods: The analytical sensitivity of LAMP and nested-PCR was obtained against10-fold serial dilutions ofT. 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 ofT. gondii DNA respectively. All the LAMP assays and nested PCRs were 100% specific. The RE-LAMP assay revealed the most sensitivity for the detection ofT. gondii DNA.Conclusions:The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection ofT. 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 ofT. gondii.
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1
Índice:
WPRIM
Tipo de estudio:
Diagnostic_studies
Idioma:
Zh
Revista:
Asian Pacific Journal of Tropical Biomedicine
Año:
2015
Tipo del documento:
Article