Development of a loop-mediated isothermal amplification assay for detection of Phytophthora sojae

fml2619-fig-0001Phytophthora sojae is a devastating pathogen that causes soybean Phytophthora root rot. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay targeting the A3aPro element for visual detection of P. sojae. The A3aPro-LAMP assay efficiently amplified the target element in < 80 min at 64 °C and was evaluated for specificity and sensitivity. The specificity was evaluated against P. sojae, Phytophthora spp., Pythium spp., and true fungi isolates. Magnesium pyrophosphate resulting from the LAMP of P. sojae could be detected by real-time measurement of turbidity. Phytophthora sojae DNA products were visualized as a ladder-like banding pattern on 2% gel electrophoresis. A positive colour (sky blue) was only observed in the presence of P. sojae with the addition of hydroxynaphthol blue prior to amplification, whereas none of other isolates showed a colour change. The detection limit of the A3aPro-specific LAMP assay for P. sojae was 10 pg μL−1 of genomic DNA per reaction. The assay also detected P. sojae from diseased soybean tissues and residues. These results suggest that the A3aPro-LAMP assay reported here can be used for the visual detection of P. sojae in plants and production fields.

The paper reports other primer pairs for an alternative PCR based detection technology, the loop-mediated isothermal amplification (LAMP) of DNA. The current database architecture does not support LAMP primers. Only the forward and backward outer primers will be loaded. Users are asked to check the original paper for primer sequences and details.

Ting-Ting Dai, Chen-Chen Lu, Jing Lu, SuoMeng Dong, WenWu Ye, YuanChao Wang, XiaoBo Zheng
FEMS Microbiology Letters
Volume 334, Issue 1, pages 27–34, September 2012

DOI: 10.1111/j.1574-6968.2012.02619.x

via Development of a loop-mediated isothermal amplification assay for detection of Phytophthora sojae – Dai – 2012 – FEMS Microbiology Letters – Wiley Online Library.


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