title: An improved validated SYBR green-based real-time quantitative PCR assay for the detection of the Penaeus stylirostris densovirus in penaeid shrimp.
abstract: The Penaeus stylirostris densovirus (PstDV) (also known as infectious hypodermal and hematopoietic necrosis virus, IHHNV), one of the major shrimp pathogens, has a worldwide distribution in farmed and wild shrimp populations. Outbreaks of IHHNV have been associated with substantial economic losses which are accompanied by a negative social impact. Current diagnostic PCR tests may result in false-positive results as several parts of PstDV genome may be endogenized in the nuclear genome of the shrimp P. stylirostris. A one-step qPCR SYBR-Green based quantitative real-time polymerase chain reaction (qPCR) assay to detect different isolates of the IHHNV in shrimp samples was developed. The detection limit of the assay was 81 viral copies of targeted DNA per reaction. The specificity of the assay was evaluated by melting curve analysis, which showed that the IHHNV product generated a single melt peak at 81.4±0.044°C. The assay was more sensitive than conventional PCR. The standardized PCR was shown to be highly sensible, specific, robust, and reproducible, which makes it an economical and powerful tool for both diagnostic applications and general research of IHHNV.
authors.affiliation: Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico.
authors.forename: Trinidad
authors.initials: T
authors.lastname: Encinas-García
authors.affiliation: Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico.
authors.forename: Fernando
authors.initials: F
authors.lastname: Mendoza-Cano
authors.affiliation: Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico.
authors.forename: Tania
authors.initials: T
authors.lastname: Enríquez-Espinoza
authors.affiliation: Departamento de Investigaciones Científicas y Tecnológicas, Universidad de Sonora, Av. Colosio s/n, entre Sahuaripa y Reforma, Hermosillo, Sonora 83000, Mexico.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Luken-Vega
authors.affiliation: Departamento de Diagnóstico Acuícola y Pesquero, Centro Nacional de Servicios de Constatación en Salud Animal (CENAPA), Carr. Fed. Cuernavaca Cuautla 8534 Colonia Progreso, Jiutepec, Morelos C.P. 62550, Mexico.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Vichido-Chávez
authors.affiliation: Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico. Electronic address: asanchez04@cibnor.mx.
authors.forename: Arturo
authors.initials: A
authors.lastname: Sánchez-Paz
mesh_terms: D000818:Animals; D017990:Densovirus / Q000302:isolation & purification*; D033561:Penaeidae / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013696:Temperature; D014773:Virology / Q000379:methods*
keywords: IHHNV; Reproducible; Sensitive; Shrimp; Specific; qPCR