Adedeji A J, Abdu P A, Luka P D, Owoade A A, Joannis T M
Viral Research Division, National Veterinary Research Institute, Vom, Nigeria.
Department of Veterinary Medicine, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria.
Vet World. 2017 Nov;10(11):1383-1388. doi: 10.14202/vetworld.2017.1383-1388. Epub 2017 Nov 26.
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.
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.
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.
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.
本研究旨在优化和应用环介导等温扩增技术（LAMP），作为传统聚合酶链反应（PCR）的替代方法，用于检测尼日利亚接种和未接种疫苗的家禽中的火鸡疱疹病毒（HVT）（FC 126株）。
使用HVT阳性对照（疫苗），利用针对该病毒HVT070基因序列的六种引物优化LAMP。这些引物可将HVT（马立克氏病病毒3型）与马立克氏病病毒1型和2型区分开来。从鸡的马立克氏病（MD）临床病例中采集样本，进行处理后进行LAMP和PCR检测。
优化了针对HVT的LAMP检测方法。通过PCR和LAMP分析的样本中，HVT的检出率分别为60%（3/5）和100%（5/5）。使用LAMP在羽毛、肝脏、皮肤和脾脏中检测到HVT，平均DNA纯度为3.05 - 4.52 μg DNA/mg（A260/A280）。传统PCR在两个接种疫苗和一个未接种疫苗的鸡样本中检测到HVT，而LAMP在两个接种疫苗和三个未接种疫苗的相应鸡样本中检测到HVT。然而，LAMP比PCR实施起来更快、更简单。
优化了用于检测HVT的LAMP检测方法。LAMP和PCR在采集的临床样本中均检测到了HVT。LAMP检测方法可作为检测HVT和其他病毒的良好替代方法，替代PCR。这是尼日利亚首次报道使用LAMP检测具有兽医重要性的病毒。LAMP应优化为尼日利亚家禽疾病（如MD）调查的诊断和研究工具。
