title: Quantitation of infectious myonecrosis virus in different tissues of naturally infected Pacific white shrimp, Litopenaeus vannamei, using real-time PCR with SYBR Green chemistry.
abstract: The Pacific white shrimp, Litopenaeus vannamei, is the most important shrimp species in volume in world aquaculture. However, in recent decades, outbreaks of diseases, especially viral diseases, have led to significant economic losses, threatening the sustainability of shrimp farming worldwide. In 2004, Brazilian shrimp farming was seriously affected by a new disease caused by the Infectious myonecrosis virus (IMNV). Thus, disease control based on rapid and sensitive pathogen detection methods has become a priority. In this study, a specific quantitation method for IMNV was developed using real-time PCR with SYBR Green chemistry and viral load of the principal target tissues of chronically infected animals was quantified. The quantitative analysis revealed that mean viral load ranged from 5.08×10(8) to 1.33×10(6)copies/μg of total RNA in the hemolymph, 5.096×10(5) to 1.26×10(3)copies/μg in the pleopods, 6.85×10(8) to 3.09×10(4)copies/μg in muscle and 8.15×10(6) to 3.90×10(3)copies/μg in gills. Different viral loads of IMNV were found with greater values in the hemolymph and muscle, followed by the pleopods and gills.
authors.affiliation: Departamento de Pesca e Aqüicultura, Universidade Federal Rural de Pernambuco, Av. Dom Manoel de Medeiros, 52171-900 Dois Irmãos, Recife, Pernambuco, Brazil.
authors.forename: Suzianny Maria Bezerra Cabral
authors.initials: SM
authors.lastname: da Silva
authors.forename: Ana Cristina de Aguiar Saldanha
authors.initials: AC
authors.lastname: Pinheiro
authors.forename: Maria Raquel Moura
authors.initials: MR
authors.lastname: Coimbra
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D001938:Brazil / Q000453:epidemiology; D003001:Cloning, Molecular; D005456:Fluorescent Dyes; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D005880:Gills / Q000821:virology; D006458:Hemolymph / Q000821:virology; D009132:Muscles / Q000821:virology; D033561:Penaeidae / Q000235:genetics / Q000821:virology*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012327:RNA Virus Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D000199:Actins; D005456:Fluorescent Dyes; D012367:RNA, Viral