Edrisi M, Varshovi H R, Safi S, Shahhoseiny M H
Department of Pathobiology, Faculty of Specialized Veterinary Sciences, Science and Research Branch, Islamic Azad University, Poonak, Hesarak, Tehran, Iran.
Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
Arch Razi Inst. 2024 Dec 31;79(6):1183-1190. doi: 10.32592/ARI.2024.79.6.1183. eCollection 2024 Dec.
Sheep pox (SP), goat pox (GP), and lumpy skin disease (LSD) are caused by subspecies of the capripox virus (CaPVs). They are significant pathogens of sheep, goats, and cattle. The causative agent is the capripox virus (CaPV), which was first isolated in South Africa. The viruses responsible for sheep pox (SP), goat pox (GP), and lumpy skin disease (LSD) are morphologically indistinguishable and have been adapted to different host species (4). Serologically, distinguishing between these viruses is challenging, and cross-immunity exists among them (2). The present study reports the evaluation and optimization of a novel loop-mediated isothermal amplification (LAMP) technique for the rapid detection of capripox viruses (CaPVs) and compares it with the polymerase chain reaction (PCR) method. LAMP primers were selected from the P32-protected gene of CaPV. The Safe-Red fluorescent dye was used to monitor the color change from red to bright yellow at a wavelength of 320 nm in positive cases, and the final results were documented through electrophoresis. The proposed LAMP test for the capripox virus demonstrated high specificity and did not cross react with other viruses in the Poxviridae family that present similar clinical symptoms. The optimized LAMP test was then compared with the PCR. The diagnostic sensitivity of LAMP and PCR was found to be identical (100%). The specificity of the LAMP test was evaluated using 30 samples of cow skin that were suspected of lumpy skin disease, along with16 additional samples, including nine positive references, fivenegative references, and two negative controls. A negative reference sample was used to assess the diagnostic sensitivity of LSDV. The proposed LAMP test is simple to implement, cost-effective, and highly sensitive, making it particularly well-suited for the detection of the capripox virus in less developed regions, laboratories, and facilities with limited resources.
绵羊痘（SP）、山羊痘（GP）和结节性皮肤病（LSD）由山羊痘病毒（CaPVs）的亚种引起。它们是绵羊、山羊和牛的重要病原体。病原体是山羊痘病毒（CaPV），该病毒最初在南非分离得到。导致绵羊痘（SP）、山羊痘（GP）和结节性皮肤病（LSD）的病毒在形态上无法区分，并且已适应不同的宿主物种（4）。在血清学上，区分这些病毒具有挑战性，并且它们之间存在交叉免疫（2）。本研究报告了一种用于快速检测山羊痘病毒（CaPVs）的新型环介导等温扩增（LAMP）技术的评估和优化，并将其与聚合酶链反应（PCR）方法进行比较。LAMP引物选自CaPV的P32保护基因。使用Safe-Red荧光染料监测阳性病例在320nm波长下从红色到亮黄色的颜色变化，最终结果通过电泳记录。所提出的用于山羊痘病毒的LAMP检测显示出高特异性，并且不会与痘病毒科中呈现相似临床症状的其他病毒发生交叉反应。然后将优化后的LAMP检测与PCR进行比较。发现LAMP和PCR的诊断敏感性相同（100%）。使用30份疑似结节性皮肤病的牛皮样本以及另外16份样本评估LAMP检测的特异性，其中包括9份阳性参考样本、5份阴性参考样本和2份阴性对照。使用阴性参考样本评估LSDV的诊断敏感性。所提出的LAMP检测易于实施、具有成本效益且高度灵敏，使其特别适合在欠发达地区、资源有限的实验室和设施中检测山羊痘病毒。
