Sensitive detection of African swine fever virus using real-time PCR with a 5' conjugated minor groove binding probe

John McKillan; Michael McMenamy; Bernt Hjertner; Francis McNeilly; Åse Uttenthal; Carmina Gallardo; Brian Adair; Gordon Allan

doi:10.1016/j.jviromet.2010.05.005

Sensitive detection of African swine fever virus using real-time PCR with a 5' conjugated minor groove binding probe

John McKillan, Michael McMenamy, Bernt Hjertner, Francis McNeilly, Åse Uttenthal, Carmina Gallardo, Brian Adair, Gordon Allan

Research output: Contribution to journal › Journal article › Research › peer-review

Abstract

The design of a 5′ conjugated minor groove binder (MGB) probe real-time PCR assay is described for the rapid, sensitive and specific detection of African swine fever virus (ASFV) DNA. The assay is designed against the 9GL region and is capable of detecting 20 copies of a DNA standard. It does not detect any of the other common swine DNA viruses tested in this study. The assay can detect ASFV DNA in a range of clinical samples. Sensitivity was equivalent to the Office International des Epizooties (OIE) recommended TaqMan assay. In addition the assay was found to have a detection limit 10-fold more sensitive than the conventional PCR recommended by the OIE. Linear range was ten logs from 2 × 101 to 2 × 1010. The assay is rapid with an amplification time just over 2 h. The development of this assay provides a useful tool for the specific diagnosis of ASF in statutory or emergency testing programs or for the detection of ASFV DNA in research applications.

Original language	English
Journal	Journal of Virological Methods
Volume	168
Issue number	1-2
Pages (from-to)	141-146
ISSN	0166-0934
DOIs	https://doi.org/10.1016/j.jviromet.2010.05.005
Publication status	E-pub ahead of print - 2010

Keywords

Real-time PCR
Minor groove binder probe
African swine fever virus

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10.1016/j.jviromet.2010.05.005

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title = "Sensitive detection of African swine fever virus using real-time PCR with a 5' conjugated minor groove binding probe",

abstract = "The design of a 5′ conjugated minor groove binder (MGB) probe real-time PCR assay is described for the rapid, sensitive and specific detection of African swine fever virus (ASFV) DNA. The assay is designed against the 9GL region and is capable of detecting 20 copies of a DNA standard. It does not detect any of the other common swine DNA viruses tested in this study. The assay can detect ASFV DNA in a range of clinical samples. Sensitivity was equivalent to the Office International des Epizooties (OIE) recommended TaqMan assay. In addition the assay was found to have a detection limit 10-fold more sensitive than the conventional PCR recommended by the OIE. Linear range was ten logs from 2 × 101 to 2 × 1010. The assay is rapid with an amplification time just over 2 h. The development of this assay provides a useful tool for the specific diagnosis of ASF in statutory or emergency testing programs or for the detection of ASFV DNA in research applications.",

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AU - McKillan, John

AU - McMenamy, Michael

AU - Hjertner, Bernt

AU - McNeilly, Francis

AU - Uttenthal, Åse

AU - Gallardo, Carmina

AU - Adair, Brian

AU - Allan, Gordon

PY - 2010

Y1 - 2010

N2 - The design of a 5′ conjugated minor groove binder (MGB) probe real-time PCR assay is described for the rapid, sensitive and specific detection of African swine fever virus (ASFV) DNA. The assay is designed against the 9GL region and is capable of detecting 20 copies of a DNA standard. It does not detect any of the other common swine DNA viruses tested in this study. The assay can detect ASFV DNA in a range of clinical samples. Sensitivity was equivalent to the Office International des Epizooties (OIE) recommended TaqMan assay. In addition the assay was found to have a detection limit 10-fold more sensitive than the conventional PCR recommended by the OIE. Linear range was ten logs from 2 × 101 to 2 × 1010. The assay is rapid with an amplification time just over 2 h. The development of this assay provides a useful tool for the specific diagnosis of ASF in statutory or emergency testing programs or for the detection of ASFV DNA in research applications.

AB - The design of a 5′ conjugated minor groove binder (MGB) probe real-time PCR assay is described for the rapid, sensitive and specific detection of African swine fever virus (ASFV) DNA. The assay is designed against the 9GL region and is capable of detecting 20 copies of a DNA standard. It does not detect any of the other common swine DNA viruses tested in this study. The assay can detect ASFV DNA in a range of clinical samples. Sensitivity was equivalent to the Office International des Epizooties (OIE) recommended TaqMan assay. In addition the assay was found to have a detection limit 10-fold more sensitive than the conventional PCR recommended by the OIE. Linear range was ten logs from 2 × 101 to 2 × 1010. The assay is rapid with an amplification time just over 2 h. The development of this assay provides a useful tool for the specific diagnosis of ASF in statutory or emergency testing programs or for the detection of ASFV DNA in research applications.

KW - Real-time PCR

KW - Minor groove binder probe

KW - African swine fever virus

U2 - 10.1016/j.jviromet.2010.05.005

DO - 10.1016/j.jviromet.2010.05.005

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JO - Journal of Virological Methods

JF - Journal of Virological Methods

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ER -

Sensitive detection of African swine fever virus using real-time PCR with a 5' conjugated minor groove binding probe

Abstract

Keywords

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