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

Dongchun Yan; Kathy F.J. Tang; Donald V. Lightner

doi:10.1016/j.jip.2009.07.008

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

Dongchun Yan, Kathy F.J. Tang, Donald V. Lightner

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

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 language	English (US)
Pages (from-to)	97-100
Number of pages	4
Journal	Journal of Invertebrate Pathology
Volume	102
Issue number	2
DOIs	https://doi.org/10.1016/j.jip.2009.07.008
State	Published - Oct 2009

Keywords

Diagnosis
Monodon baculovirus
Real-time PCR
Sensitivity
Specificity

ASJC Scopus subject areas

Ecology, Evolution, Behavior and Systematics

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10.1016/j.jip.2009.07.008

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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 Tang, {Kathy F.J.} and Lightner, {Donald V.}",

note = "Funding Information: This work was funded by the International Cooperate Foundation for Excellent University Teacher in Shandong Province (2007), a Project of Shandong Province Higher Educational Science and Technology Program (2009), and a Grant from USDA, CSREES, to the US Marine Shrimp Farming Consortium.",

year = "2009",

month = oct,

doi = "10.1016/j.jip.2009.07.008",

language = "English (US)",

volume = "102",

pages = "97--100",

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AU - Tang, Kathy F.J.

AU - Lightner, Donald V.

N1 - Funding Information: This work was funded by the International Cooperate Foundation for Excellent University Teacher in Shandong Province (2007), a Project of Shandong Province Higher Educational Science and Technology Program (2009), and a Grant from USDA, CSREES, to the US Marine Shrimp Farming Consortium.

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

Abstract

Keywords

ASJC Scopus subject areas

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