Development of a real-time PCR assay for detection of monodon baculovirus (MBV) in penaeid shrimp

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Abstract

A real-time PCR method was developed to detect monodon baculovirus (MBV) in penaeid shrimp. A pair of MBV primers to amplify a 135 bp DNA fragment and a TaqMan probe were developed. The primers and TaqMan probe were specific for MBV and did not cross react with Hepatopancreatic parvovirus (HPV), White spot syndrome virus (WSSV), Infectious hypodermal and haematopoietic virus (IHHNV) and specific pathogen free (SPF) shrimp DNA. A plasmid (pMBV) containing the target MBV sequence was constructed and used for determination of the sensitivity of the real-time PCR. This real-time PCR assay had a detection limit of one plasmid MBV DNA copy. Most significantly, this real-time PCR method can detect MBV positive samples from different geographic locations in the University of Arizona collection, including Thailand and Indonesia collected over a 13-year period.

Original languageEnglish (US)
Pages (from-to)97-100
Number of pages4
JournalJournal of Invertebrate Pathology
Volume102
Issue number2
DOIs
StatePublished - Oct 2009

Fingerprint

Monodon
Penaeidae
Baculoviridae
quantitative polymerase chain reaction
assay
assays
plasmid
DNA
virus
probe
plasmids
White spot syndrome virus
Protoparvovirus
pathogen
Thailand
Indonesia
detection
detection limit
shrimp
viruses

Keywords

  • Diagnosis
  • Monodon baculovirus
  • Real-time PCR
  • Sensitivity
  • Specificity

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics

Cite this

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title = "Development of a real-time PCR assay for detection of monodon baculovirus (MBV) in penaeid shrimp",
abstract = "A real-time PCR method was developed to detect monodon baculovirus (MBV) in penaeid shrimp. A pair of MBV primers to amplify a 135 bp DNA fragment and a TaqMan probe were developed. The primers and TaqMan probe were specific for MBV and did not cross react with Hepatopancreatic parvovirus (HPV), White spot syndrome virus (WSSV), Infectious hypodermal and haematopoietic virus (IHHNV) and specific pathogen free (SPF) shrimp DNA. A plasmid (pMBV) containing the target MBV sequence was constructed and used for determination of the sensitivity of the real-time PCR. This real-time PCR assay had a detection limit of one plasmid MBV DNA copy. Most significantly, this real-time PCR method can detect MBV positive samples from different geographic locations in the University of Arizona collection, including Thailand and Indonesia collected over a 13-year period.",
keywords = "Diagnosis, Monodon baculovirus, Real-time PCR, Sensitivity, Specificity",
author = "Dongchun Yan and Feng-Jyu Tang-Nelson and Lightner, {Donald V}",
year = "2009",
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language = "English (US)",
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T1 - Development of a real-time PCR assay for detection of monodon baculovirus (MBV) in penaeid shrimp

AU - Yan, Dongchun

AU - Tang-Nelson, Feng-Jyu

AU - Lightner, Donald V

PY - 2009/10

Y1 - 2009/10

N2 - A real-time PCR method was developed to detect monodon baculovirus (MBV) in penaeid shrimp. A pair of MBV primers to amplify a 135 bp DNA fragment and a TaqMan probe were developed. The primers and TaqMan probe were specific for MBV and did not cross react with Hepatopancreatic parvovirus (HPV), White spot syndrome virus (WSSV), Infectious hypodermal and haematopoietic virus (IHHNV) and specific pathogen free (SPF) shrimp DNA. A plasmid (pMBV) containing the target MBV sequence was constructed and used for determination of the sensitivity of the real-time PCR. This real-time PCR assay had a detection limit of one plasmid MBV DNA copy. Most significantly, this real-time PCR method can detect MBV positive samples from different geographic locations in the University of Arizona collection, including Thailand and Indonesia collected over a 13-year period.

AB - A real-time PCR method was developed to detect monodon baculovirus (MBV) in penaeid shrimp. A pair of MBV primers to amplify a 135 bp DNA fragment and a TaqMan probe were developed. The primers and TaqMan probe were specific for MBV and did not cross react with Hepatopancreatic parvovirus (HPV), White spot syndrome virus (WSSV), Infectious hypodermal and haematopoietic virus (IHHNV) and specific pathogen free (SPF) shrimp DNA. A plasmid (pMBV) containing the target MBV sequence was constructed and used for determination of the sensitivity of the real-time PCR. This real-time PCR assay had a detection limit of one plasmid MBV DNA copy. Most significantly, this real-time PCR method can detect MBV positive samples from different geographic locations in the University of Arizona collection, including Thailand and Indonesia collected over a 13-year period.

KW - Diagnosis

KW - Monodon baculovirus

KW - Real-time PCR

KW - Sensitivity

KW - Specificity

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DO - 10.1016/j.jip.2009.07.008

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VL - 102

SP - 97

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JO - Journal of Invertebrate Pathology

JF - Journal of Invertebrate Pathology

SN - 0022-2011

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ER -