On-site detection of bovine leukemia virus by a field-deployable automatic nucleic extraction plus insulated isothermal polymerase chain reaction system.

Abstract

Bovine leukemia virus (BLV) is a contagious, oncogenic deltaretrovirus of cattle with a worldwide distribution. In the US, over 40% of dairy cows are infected with the virus, and evidence of its economic impact is growing. This study evaluated the performance of a field-deployable automatic nucleic acid-extraction/insulated isothermal PCR (iiPCR) system for on-site BLV-proviral DNA detection in dairy cows compared with a conventional laboratory real-time PCR (rt-PCR). Assay performance was verified in parallel tests of 36 archived blood samples with 100% agreement (κ = 1.0; n = 36) between the iiPCR and conventional rt-PCR systems, and the limit of detection of the iiPCR assay was estimated to be 4 copies (genome equivalent) per reaction. The field-deployable iiPCR system was subsequently used on-farm to test freshly collected blood samples, and showed 100% agreement (κ = 1.0; n = 32) with the laboratory rt-PCR system. Fresh blood samples were collected on a second farm and tested on both systems, also with 100% agreement (κ = 1.0; n = 34). The field-deployable iiPCR/POCKIT™ combo system performs as well as a conventional laboratory-based rt-PCR system for detection of BLV proviral DNA in whole blood and may be a useful tool for on-farm evaluation of BLV-infection status in dairy cattle.

• Authors: Vickie J Ruggiero, Oscar J Benitez, Yun-Long Tsai, Pei-Yu Alison Lee, Chuan-Fu Tsai, Yu-Chun Lin, Hsiao-Fen Grace Chang, Hwa-Tang Thomas Wang, Paul Bartlett
• Journal: Journal of virological methods (J Virol Methods) Vol. 259 Pg. 116-121 (09 2018) ISSN: 1879-0984 [Electronic] Netherlands
• PMID: 29902491 (Publication Type: Evaluation Study, Journal Article, Research Support, U.S. Gov't, Non-P.H.S.)
• Copyright: Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Chemical References

• DNA, Viral

Topics

• Animals
• Automation (methods)
• Cattle
• DNA, Viral (genetics, isolation & purification)
• Enzootic Bovine Leukosis (diagnosis, virology)
• Leukemia Virus, Bovine (genetics, isolation & purification)
• Molecular Diagnostic Techniques (methods)
• Point-of-Care Testing
• Polymerase Chain Reaction (methods)
• Proviruses (genetics, isolation & purification)