Henan Provincial Engineering Laboratory of Insects Bio-reactor, China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, Nanyang, 473061, PR China.
Henan Provincial Engineering Laboratory of Insects Bio-reactor, China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, Nanyang, 473061, PR China.
Poult Sci. 2020 Feb;99(2):791-796. doi: 10.1016/j.psj.2019.12.024. Epub 2020 Jan 24.
In this study, loop-mediated isothermal amplification (LAMP) was used to establish a rapid, specific, and visual detection method for duck hepatitis B virus (DHBV). The design and synthesis of 4 specific LAMP primers were based on the conserved gene region of the DHBV genome, and the optimum temperature and time of the LAMP reaction were 63°C and 50 min, respectively. The LAMP assay was confirmed to be specific for DHBV detection and had the same sensitivity as the quantitative PCR assay. A visual detection method for rapid determination of results was developed using a color indicator containing phenol red and cresol red. A color change was produced based on a pH change in the reaction system, indicating a positive reaction. For the detection of samples from ducks and geese, the LAMP method has the advantages of simplicity, high sensitivity and specificity, good visibility, and low cost. Moreover, it is more practical and convenient than PCR-related assays for the clinical detection of DHBV.
在这项研究中，我们使用环介导等温扩增（LAMP）技术建立了一种快速、特异、可视化的鸭乙型肝炎病毒（DHBV）检测方法。根据 DHBV 基因组保守区设计并合成了 4 条特异性 LAMP 引物，LAMP 反应的最适温度和时间分别为 63°C 和 50 min。该 LAMP 检测方法特异性好，与定量 PCR 检测方法具有相同的灵敏度。我们还开发了一种使用含有苯酚红和甲酚红的显色指示剂的可视化检测方法，用于快速确定结果。根据反应体系中 pH 值的变化，显色指示剂产生颜色变化，指示阳性反应。对于鸭和鹅样本的检测，LAMP 方法具有操作简单、灵敏度和特异性高、可视性好、成本低等优点，而且比与 PCR 相关的检测方法更适合 DHBV 的临床检测。
