Development of PCR Diagnosis for Shrimp Acute Hepatopancreatic Necrosis Disease (AHPND) Strain of Vibrio parahaemolyticus

Necrosis aguda del hepatopáncreas: una revisión de la enfermedad en Penaeus vannamei

“…El impacto generado por las patologías bacterianas en la camaronicultura no es reciente, históricamente las infecciones causadas por bacterias, en especial por las especies pertenecientes al género Vibrio, han sido responsables de altas mortalidades (Martin et al, 2004;Morales-Covarrubias, 2004;Aguirre-Guzmán et al, 2005;Trujillo et al, 2005;Soto et al, 2010;Peña y Varela, 2013;Tinwongger et al, 2014;Varela y Peña, 2016). Algunas de las especies reportadas incluyen a V. parahaemolyticus, V. penaeicida, V. nigripulchritudo, V. alginolyticus, V. harveyi y V. campbelli (Morales-Covarrubias, 2004;Soto et al, 2010; Morales-Covarrubias y Gómez-Gil, 2014), con grados de afectación y virulencia variables.…”

Detection of acute hepatopancreatic necrosis disease strain of Vibrio parahaemolyticus using loop‐mediated isothermal amplification

“…parahaemolyticus (TUMSAT‐Vp3) was recently reported (Tinwongger et al . ). However, PCR takes several hours and requires an expensive thermal cycler.…”

“…We used six V. parahaemolyticus strains isolated from shrimp farms in Thailand (Tinwongger et al . ): two AHPND (E2 and D6) strains and four non‐AHPND (N7, N10, FP11 and FP14) strains. Total genomic DNA was extracted by the CTAB method (Sambrook & Russel ).…”

A rapid method for simultaneously diagnosing four shrimp diseases using PCR‐DNA chromatography method

“…Total genomic DNA of these samples was extracted using taco™ Nucleic Acid Automatic Extraction System (GeneReach Biotechnology Corp, Taichung City, Taiwan). For each of the DNA samples, each disease was detected by conventional PCR using KAPA2G Fast Multiplex Mix (Kapa Biosystems, Wilmington, MA, USA) or KAPA 2G Fast Hot Start Ready Mix with dye (Kapa Biosystems) referring to previous reports: WSD (Lo et al., ), IHHN (Tang, Navarro, & Lightner, ), AHPND (Tinwongger et al., ) and EHP infections (Tangprasittipap et al., ). We obtained 30 samples that were positive for white spot virus (WSV) which is the causative virus of WSD, seven samples that were positive for infectious hypodermal and haematopoietic necrosis virus (IHHNV) which is the causative virus of IHHN, 14 samples that were positive for toxin gene of a virulent strain of Vibrio parahaemolyticus of AHPND, 19 samples that were positive for EHP, 14 samples that were double‐positive for IHHNV and EHP and samples that were negative for each of these diseases.…”

“…To develop PCR‐DNA chromatography, we targeted WSD, IHHN, AHPND and EHP infections and 16S rRNA/tRNA Val /12S rRNA mitochondrial region (an internal control) using primers previously reported for WSD (Kiatpathomchai et al., ), IHHN (Tang et al., ), AHPND (Tinwongger et al., ), EHP infections (Tangprasittipap et al., ) and 16S rRNA/tRNA Val /12S rRNA (Pascoal, Barros‐Velázquez, Cepeda, Gallardo, & Calo‐Mata, ) (Table ). The primers for IHHN and AHPND are also recommended by the World Organisation for Animal Health ().…”