Detection of infectious myonecrosis virus (IMNV) of penaeid shrimp by reverse-transcriptase polymerase chain reaction (RT-PCR)

Bonnie T. Poulos, Donald V Lightner

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Infectious myonecrosis virus (IMNV) infecting cultured Litopenaeus vannamei in Brazil is & double-stranded RNA virus that causes a slowly progressive disease with cumulative mortalities of up to 70%. The disease is currently diagnosed using a combination of gross signs (primarily skeletal tail muscle necrosis with white opaque discoloration), histopathology, and in situ hybridization with a digoxigenin-labeled gene probe. A rapid and sensitive method for definitive diagnosis of the disease was developed using reverse-transcriptase polymerase chain reaction (RT-PCR). Two primer sets were used to detect 328 and 139 bp amplicons in a nested RT-PCR assay. Using RNA extracted from purified virions, the first step reaction detected 100 copies of the IMNV viral genome whereas the nested step detected 10 copies. The primers were shown to be specific for IMNV and no amplicons were detected using RNA extracted from shrimp infected with other penaeid shrimp viruses (Taura syndrome virus [TSV], yellowhead virus [YHV], infectious hypodermal hematopoietic necrosis virus [IHHNV] and white spot syndrome virus [WSSV]).

Original languageEnglish (US)
Pages (from-to)69-72
Number of pages4
JournalDiseases of Aquatic Organisms
Volume73
Issue number1
StatePublished - Nov 21 2006

Fingerprint

Penaeidae
polymerase chain reaction
virus
reverse transcriptase polymerase chain reaction
viruses
Taura syndrome virus
dsRNA viruses
RNA
Infectious hematopoietic necrosis virus
digoxigenin
White spot syndrome virus
Litopenaeus vannamei
disease diagnosis
discoloration
virion
histopathology
in situ hybridization
necrosis
shrimp
tail

Keywords

  • IMNV
  • Infectious myonecrosis
  • Penaeid shrimp
  • RT-PCR

ASJC Scopus subject areas

  • Aquatic Science
  • Ecology

Cite this

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abstract = "Infectious myonecrosis virus (IMNV) infecting cultured Litopenaeus vannamei in Brazil is & double-stranded RNA virus that causes a slowly progressive disease with cumulative mortalities of up to 70{\%}. The disease is currently diagnosed using a combination of gross signs (primarily skeletal tail muscle necrosis with white opaque discoloration), histopathology, and in situ hybridization with a digoxigenin-labeled gene probe. A rapid and sensitive method for definitive diagnosis of the disease was developed using reverse-transcriptase polymerase chain reaction (RT-PCR). Two primer sets were used to detect 328 and 139 bp amplicons in a nested RT-PCR assay. Using RNA extracted from purified virions, the first step reaction detected 100 copies of the IMNV viral genome whereas the nested step detected 10 copies. The primers were shown to be specific for IMNV and no amplicons were detected using RNA extracted from shrimp infected with other penaeid shrimp viruses (Taura syndrome virus [TSV], yellowhead virus [YHV], infectious hypodermal hematopoietic necrosis virus [IHHNV] and white spot syndrome virus [WSSV]).",
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AB - Infectious myonecrosis virus (IMNV) infecting cultured Litopenaeus vannamei in Brazil is & double-stranded RNA virus that causes a slowly progressive disease with cumulative mortalities of up to 70%. The disease is currently diagnosed using a combination of gross signs (primarily skeletal tail muscle necrosis with white opaque discoloration), histopathology, and in situ hybridization with a digoxigenin-labeled gene probe. A rapid and sensitive method for definitive diagnosis of the disease was developed using reverse-transcriptase polymerase chain reaction (RT-PCR). Two primer sets were used to detect 328 and 139 bp amplicons in a nested RT-PCR assay. Using RNA extracted from purified virions, the first step reaction detected 100 copies of the IMNV viral genome whereas the nested step detected 10 copies. The primers were shown to be specific for IMNV and no amplicons were detected using RNA extracted from shrimp infected with other penaeid shrimp viruses (Taura syndrome virus [TSV], yellowhead virus [YHV], infectious hypodermal hematopoietic necrosis virus [IHHNV] and white spot syndrome virus [WSSV]).

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