Establishment of universal loop-mediated isothermal amplification method (LAMP) for rapid detection of pathogenic Vibrio spp. in aquatic organisms

Xu, Haisheng; He, Lin; Lv, Sunjian; Gong, Qi; Li, Song

doi:10.5897/ajmr11.1417

Cited by 2 publications

(2 citation statements)

References 47 publications

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“…El método LAMP se ha utilizado para la detección de patógenos de mariscos en muchos países (Kono et al 2004, Mekata et al 2006, Yamazaki et al 2008, Jaroenram et al 2009, Mekata et al 2009, Chen y Ge 2010, Arunrut et al 2011, Maralit et al 2012, Xu et al 2012, Feng et al 2013, Kongrueng et al 2015. Diversos informes muestran inconsistencias en los protocolos LAMP para detectar patógenos de diferentes ubicaciones geográficas.…”

Section: Positive Samplesunclassified

“…LAMP has been used to detect shellfish pathogens in many countries (Kono et al 2004, Mekata et al 2006, Yamazaki et al 2008, Jaroenram et al 2009, Mekata et al 2009, Chen and Ge 2010, Arunrut et al 2011, Maralit et al 2012, Xu et al 2012, Feng et al 2013, Kongrueng et al 2015. Many reports show inconsistencies with LAMP protocols for detecting pathogens from different geographic locations.…”

Section: Introductionmentioning

confidence: 99%

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Loop-mediated isothermal amplification for diagnosing marine pathogens in tissues of Crassostrea spp. and white shrimp, Litopenaeus vannamei, farmed in Mexico

Mendoza-Avilés¹,

Muñoz-Rojas

Rojas

et al. 2021

Cienc. Mar.

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Loop-mediated isothermal amplification (LAMP) is an accurate, sensitive, rapid, and easy-to-perform method for gene amplification under isothermal conditions, and it has served as a powerful diagnostic tool. In this study, we used LAMP to develop a diagnostic protocol for detecting Vibrio parahaemolyticus and white spot syndrome virus in whiteleg shrimp, and Perkinsus spp. in Crassostrea spp. in Mexico. These pathogens are associated with different diseases and are considered a threat in the aquaculture industry. Infected and uninfected oysters and shrimp were obtained from farms in the northwest coast of Mexico to standardize the LAMP assay. We determined the candidate target genes in the first-round analysis of many sets of primers, and then we chose a set of primers that successfully amplified with Mexican samples. We optimized the LAMP reactions for each pathogen with the chosen primer sets using temperature gradients from 61 to 65 ºC, DNA concentrations from 2.5 pg to 250.0 ng, and reaction times from 10 to 60 min. This study established a diagnostic procedure for detecting pathogens in oysters and shrimp from Mexico. Early diagnosis and treatment of pathogens can immensely reduce disease transmission in aquaculture farms.

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