Abstract
A nucleic acid-based multiplexed assay was developed that combines detection of foot-and-mouth disease virus (FMDV) with rule-out assays for two other foreign animal diseases and four domestic animal diseases that cause vesicular or ulcerative lesions indistinguishable from FMDV infection in cattle, sheep and swine. The FMDV "look-alike" diagnostic assay panel contains 5 PCR and 12 reverse transcriptase PCR (RT-PCR) signatures for a total of 17 simultaneous PCR amplifications for 7 diseases plus incorporating 4 internal assay controls. It was developed and optimized to amplify both DNA and RNA viruses simultaneously in a single tube and employs Luminex™ liquid array technology. Assay development including selection of appropriate controls, a comparison of signature performance in single and multiplex testing against target nucleic acids, as well of limits of detection for each of the individual signatures is presented. While this assay is a prototype and by no means a comprehensive test for FMDV "look-alike" viruses, an assay of this type is envisioned to have benefit to a laboratory network in routine surveillance and possibly for post-outbreak proof of freedom from foot-and-mouth disease.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 61-69 |
| Number of pages | 9 |
| Journal | Journal of Virological Methods |
| Volume | 153 |
| Issue number | 1 |
| DOIs | |
| State | Published - Oct 2008 |
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Keywords
- FMDV
- Multiplex
- RT-PCR
- Rule-out
ASJC Scopus subject areas
- Virology
Cite this
Multiplexed molecular assay for rapid exclusion of foot-and-mouth disease. / Lenhoff, Raymond J.; Naraghi-Arani, Pejman; Thissen, James B.; Olivas, Jason; Carillo, A. Celena; Chinn, Corey; Rasmussen, Max; Messenger, Sharon M.; Suer, Lynn D.; Smith, Sally M.; Tammero, Lance F Bentley; Vitalis, Elizabeth A.; Slezak, Thomas R.; Hullinger, Pam; Hindson, Benjamin J.; Hietala, Sharon K.; Crossley, Beate; McBride, Mary T.
In: Journal of Virological Methods, Vol. 153, No. 1, 10.2008, p. 61-69.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Multiplexed molecular assay for rapid exclusion of foot-and-mouth disease
AU - Lenhoff, Raymond J.
AU - Naraghi-Arani, Pejman
AU - Thissen, James B.
AU - Olivas, Jason
AU - Carillo, A. Celena
AU - Chinn, Corey
AU - Rasmussen, Max
AU - Messenger, Sharon M.
AU - Suer, Lynn D.
AU - Smith, Sally M.
AU - Tammero, Lance F Bentley
AU - Vitalis, Elizabeth A.
AU - Slezak, Thomas R.
AU - Hullinger, Pam
AU - Hindson, Benjamin J.
AU - Hietala, Sharon K.
AU - Crossley, Beate
AU - McBride, Mary T.
PY - 2008/10
Y1 - 2008/10
N2 - A nucleic acid-based multiplexed assay was developed that combines detection of foot-and-mouth disease virus (FMDV) with rule-out assays for two other foreign animal diseases and four domestic animal diseases that cause vesicular or ulcerative lesions indistinguishable from FMDV infection in cattle, sheep and swine. The FMDV "look-alike" diagnostic assay panel contains 5 PCR and 12 reverse transcriptase PCR (RT-PCR) signatures for a total of 17 simultaneous PCR amplifications for 7 diseases plus incorporating 4 internal assay controls. It was developed and optimized to amplify both DNA and RNA viruses simultaneously in a single tube and employs Luminex™ liquid array technology. Assay development including selection of appropriate controls, a comparison of signature performance in single and multiplex testing against target nucleic acids, as well of limits of detection for each of the individual signatures is presented. While this assay is a prototype and by no means a comprehensive test for FMDV "look-alike" viruses, an assay of this type is envisioned to have benefit to a laboratory network in routine surveillance and possibly for post-outbreak proof of freedom from foot-and-mouth disease.
AB - A nucleic acid-based multiplexed assay was developed that combines detection of foot-and-mouth disease virus (FMDV) with rule-out assays for two other foreign animal diseases and four domestic animal diseases that cause vesicular or ulcerative lesions indistinguishable from FMDV infection in cattle, sheep and swine. The FMDV "look-alike" diagnostic assay panel contains 5 PCR and 12 reverse transcriptase PCR (RT-PCR) signatures for a total of 17 simultaneous PCR amplifications for 7 diseases plus incorporating 4 internal assay controls. It was developed and optimized to amplify both DNA and RNA viruses simultaneously in a single tube and employs Luminex™ liquid array technology. Assay development including selection of appropriate controls, a comparison of signature performance in single and multiplex testing against target nucleic acids, as well of limits of detection for each of the individual signatures is presented. While this assay is a prototype and by no means a comprehensive test for FMDV "look-alike" viruses, an assay of this type is envisioned to have benefit to a laboratory network in routine surveillance and possibly for post-outbreak proof of freedom from foot-and-mouth disease.
KW - FMDV
KW - Multiplex
KW - RT-PCR
KW - Rule-out
UR - http://www.scopus.com/inward/record.url?scp=49349115293&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=49349115293&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2008.06.015
DO - 10.1016/j.jviromet.2008.06.015
M3 - Article
VL - 153
SP - 61
EP - 69
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -