Avian Disease Division, Animal and Plant Quarantine Agency, HyukSin 8-ro, GimCheon, Republic of Korea.
Avian Disease Division, Animal and Plant Quarantine Agency, HyukSin 8-ro, GimCheon, Republic of Korea.
J Virol Methods. 2018 Jun;256:6-11. doi: 10.1016/j.jviromet.2018.02.013. Epub 2018 Feb 21.
Loop-mediated isothermal amplification (LAMP) methods to detect chicken infectious anemia virus (CIAV), reticuloendotheliosis virus (REV), and Marek's disease virus (MDV), and a reverse transcription (RT)-LAMP assay to detect infectious bursal disease virus (IBDV), were developed. The CIAV-LAMP, REV-LAMP, MDV-LAMP, and IBDV-RT-LAMP methods were performed using four sets of six primers targeting the VP1 gene of CIAV, the gp90 gene of REV, the Meq gene of MDV, and the VP2 gene of IBDV. The results (a change in color) were observed visually. The methods showed high specificity and sensitivity. The detection limits were 50 genomic copies of CIAV, 16 genomic copies of REV, 20 genomic copies of MDV, and 250 genomic copies of IBDV. When used to test clinical samples, the results of the LAMP assays were in 100% agreement with a previously described PCR. Therefore, the LAMP assays are simple, rapid, highly sensitive, and specific methods for detecting four immune-suppressive viruses.
建立了用于检测鸡传染性贫血病毒(CIAV)、网状内皮组织增生症病毒(REV)和马立克氏病病毒(MDV)的环介导等温扩增(LAMP)方法,以及一种用于检测传染性法氏囊病病毒(IBDV)的逆转录-LAMP 检测方法。CIAV-LAMP、REV-LAMP、MDV-LAMP 和 IBDV-RT-LAMP 方法使用针对 CIAV 的 VP1 基因、REV 的 gp90 基因、MDV 的 Meq 基因和 IBDV 的 VP2 基因的四组六对引物进行。结果(颜色变化)通过肉眼观察。这些方法具有高度的特异性和敏感性。检测限分别为 CIAV 的 50 个基因组拷贝、REV 的 16 个基因组拷贝、MDV 的 20 个基因组拷贝和 IBDV 的 250 个基因组拷贝。当用于检测临床样本时,LAMP 检测方法的结果与先前描述的 PCR 方法完全一致。因此,LAMP 检测方法是检测四种免疫抑制病毒的简单、快速、高灵敏度和特异性方法。