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Application of Loop-Mediated Isothermal Amplification Assay in the Detection of Herpesvirus of Turkey (FC 126 Strain) From Chicken Samples in Nigeria


Affiliations
1 Viral Research Division, National Veterinary Research Institute, Vom, Nigeria
2 Department of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria
3 Biotechnology Division, National Veterinary Research Institute, Vom, Nigeria
4 Department of Veterinary Medicine, University of Ibadan, Nigeria
5 Regional Laboratory for Animal Influenza and Other Transboundary Animal Diseases, National Veterinary Research Institute, Vom, Nigeria
 

Aim: This study was designed to optimize and apply the use of loop-mediated isothermal amplification (LAMP) as an alternative to conventional polymerase chain reaction (PCR) for the detection of herpesvirus of turkeys (HVT) (FC 126 strain) in vaccinated and non-vaccinated poultry in Nigeria.
Materials and Methods: HVT positive control (vaccine) was used for optimization of LAMP using six primers that target the HVT070 gene sequence of the virus. These primers can differentiate HVT, a Marek’s disease virus (MDV) serotype 3 from MDV serotypes 1 and 2. Samples were collected from clinical cases of Marek’s disease (MD) in chickens, processed and subjected to LAMP and PCR.
Results: LAMP assay for HVT was optimized. HVT was detected in 60% (3/5) and 100% (5/5) of the samples analyzed by PCR and LAMP, respectively. HVT was detected in the feathers, liver, skin, and spleen with average DNA purity of 3.05-4.52 μg DNA/mg (A260/A280) using LAMP. Conventional PCR detected HVT in two vaccinated and one unvaccinated chicken samples, while LAMP detected HVT in two vaccinated and three unvaccinated corresponding chicken samples. However, LAMP was a faster and simpler technique to carry out than PCR.
Conclusion: LAMP assay for the detection of HVT was optimized. LAMP and PCR detected HVT in clinical samples collected. LAMP assay can be a very good alternative to PCR for detection of HVT and other viruses. This is the first report of the use of LAMP for the detection of viruses of veterinary importance in Nigeria. LAMP should be optimized as a diagnostic and research tool for investigation of poultry diseases such as MD in Nigeria.

Keywords

Herpesvirus of Turkeys, Loop-Mediated Isothermal Amplification Procedure, Nigeria.
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  • Application of Loop-Mediated Isothermal Amplification Assay in the Detection of Herpesvirus of Turkey (FC 126 Strain) From Chicken Samples in Nigeria

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Authors

A. J. Adedeji
Viral Research Division, National Veterinary Research Institute, Vom, Nigeria
P. A. Abdu
Department of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria
P. D. Luka
Biotechnology Division, National Veterinary Research Institute, Vom, Nigeria
A. A. Owoade
Department of Veterinary Medicine, University of Ibadan, Nigeria
T. M. Joannis
Regional Laboratory for Animal Influenza and Other Transboundary Animal Diseases, National Veterinary Research Institute, Vom, Nigeria

Abstract


Aim: This study was designed to optimize and apply the use of loop-mediated isothermal amplification (LAMP) as an alternative to conventional polymerase chain reaction (PCR) for the detection of herpesvirus of turkeys (HVT) (FC 126 strain) in vaccinated and non-vaccinated poultry in Nigeria.
Materials and Methods: HVT positive control (vaccine) was used for optimization of LAMP using six primers that target the HVT070 gene sequence of the virus. These primers can differentiate HVT, a Marek’s disease virus (MDV) serotype 3 from MDV serotypes 1 and 2. Samples were collected from clinical cases of Marek’s disease (MD) in chickens, processed and subjected to LAMP and PCR.
Results: LAMP assay for HVT was optimized. HVT was detected in 60% (3/5) and 100% (5/5) of the samples analyzed by PCR and LAMP, respectively. HVT was detected in the feathers, liver, skin, and spleen with average DNA purity of 3.05-4.52 μg DNA/mg (A260/A280) using LAMP. Conventional PCR detected HVT in two vaccinated and one unvaccinated chicken samples, while LAMP detected HVT in two vaccinated and three unvaccinated corresponding chicken samples. However, LAMP was a faster and simpler technique to carry out than PCR.
Conclusion: LAMP assay for the detection of HVT was optimized. LAMP and PCR detected HVT in clinical samples collected. LAMP assay can be a very good alternative to PCR for detection of HVT and other viruses. This is the first report of the use of LAMP for the detection of viruses of veterinary importance in Nigeria. LAMP should be optimized as a diagnostic and research tool for investigation of poultry diseases such as MD in Nigeria.

Keywords


Herpesvirus of Turkeys, Loop-Mediated Isothermal Amplification Procedure, Nigeria.