Validation of a reference control for an SYBR-Green fluorescence assay-based real-time PCR for detection of bovine herpesvirus 5 in experimentally exposed bovine embryos

The objective of this study was to optimize an internal control to improve SYBR-Green-based qPCR to amplify/detect the BoHV-5 US9 gene in bovine embryos produced invitro and experimentally exposed to the virus. We designed an SYBR-Green-based binding assay that is quick to perform, reliable, easily optimized and compares well with the published assay. Herein we demonstrated its general applicability to detect BoHV-5 US9 gene in bovine embryos produced invitro experimentally exposed to BoHV-5. In order to validate the assay, three different reference genes were tested; and the histone 2a gene was shown to be the most adequate for normalizing the qPCR reaction, by considering melting and standard curves ( p<0.05). On the other hand, no differences were found in the development of bovine embryos invitro whether they were exposed to BoHV-5 reference and field strains comparing to unexposed embryos. The developed qPCR assay may have important field applications as it provides an accurate BoHV-5 US9 gene detection using a proven reference gene and is considerably less expensive than the TaqMan qPCR currently employed in sanitary programs. © 2013 Elsevier Ltd.

Energy resources development opportunities in developing countries, with special reference to Latin America

Includes bibliography

The sisterhood method to estimate maternal mortality: applications with special reference to Latin America

Includes bibliography

Considerations governing the selection and design of waterwheel generators with special reference to the conditions in Latin America

Includes bibliography

Some considerations on the nature of the task faced by the IRG, and a few suggestions as to how to organize its activities, with particular reference to Latin America

Includes bibliography

Some considerations on the nature of the task faced by the IRG, and a few suggestions as to how to organize its activities, with particular reference to Latin America

Includes bibliography

Review of the development of the transport system in the Caribbean with reference to the establishment of regional institutions and the involvement of aid donors

Includes bibliography

Development and legal aspects of general insurance in the Caribbean with special reference to the OECS countries

Examina y analiza los principales asuntos legales relacionados al desarrollo y operación de las compañías de seguros generales en el Caribe, con especial referencia a los países de la Organización de Estados del Caribe Oriental.

Countertrade policies and practices with special reference to selected Caribbean countries

Includes bibliography