The distribution and frequency of potato leaf roll virus in the four most important potato growing regions in Serbia were studied during the seven years (2012–18). One hundred randomly collected potato tubers were sampled from each seed lot. The young leaves that developed in three weeks were sampled and tested to record infection rate. The presence of potato leaf roll virus was detected by double-antibody sandwich enzyme-linked immunosorbent assay and disease incidence was calculated using standard formula. The obtained result showed that the highest prevalence of potato leaf roll virus was detected from seed potato samples originated from the Raški region during 2018 (20.7%), while in the Moravički region, only 2.3–11.1% of the potato leaf roll virus was detected every year. The average annual potato leaf roll virus infection was the highest in 2012 (8.4%) and 2018 (8.0%). For further confirmation of potato leaf roll virus infection, reverse transcription (RT)-PCR was performed using specific primers PLRVCPvEcoRI /PLRVCPcNcoI, designed to amplify a 650 bp fragment of the full-length coat protein gene. The PCR products derived from 26 isolates were directly sequenced using the same primer pair as in RT-PCR. The coat protein sequence analysis revealed that the Serbian potato leaf roll virus isolates showed very low nucleotide diversity (95.9–100%). They shared the highest nt identities of 98.08–99.36% with the sequences of potato leaf roll virus isolates deposited in the GenBank from other parts of the world. Phylogenetic analysis and the haplotype network of the coat protein gene sequences showed that the Serbian potato leaf roll virus isolates could be classified in two different groups indicating two possible introductions of the virus to Serbia. The results of this study confirmed the importance of potato leaf roll virus in seed potato production in Serbia. Additionally, this research highlights the need for a continuous monitoring of the potato seeds produced in Serbia as well as imported seeds for the presence of potato leaf roll virus.

在这七年中（2012-18年），对塞尔维亚四个最重要的马铃薯种植区中马铃薯叶卷病毒的分布和频率进行了研究。从每个种子批次中随机抽取一百个马铃薯块茎。对三周内发育的幼叶进行采样并测试以记录感染率。通过双抗体夹心酶联免疫吸附法检测马铃薯叶卷病毒的存在，并使用标准公式计算疾病发病率。获得的结果表明，在2018年期间，从Raški地区的种子马铃薯样品中检出的马铃薯叶卷病毒的发病率最高（20.7％），而在Moravički地区，检出的马铃薯叶卷病毒仅为2.3–11.1％每年。马铃薯叶卷病毒的年平均感染率在2012年（8.4％）和2018年（8.0％）最高。为了进一步确认马铃薯叶卷病毒感染，使用特异性引物PLRVCPv进行了逆转录（RT）-PCREco RI / PLRVCPc NcoI，旨在扩增全长外壳蛋白基因的650 bp片段。使用与RT-PCR中相同的引物对，对来自26个分离株的PCR产物进行直接测序。外壳蛋白序列分析表明，塞尔维亚马铃薯卷叶病毒分离株显示出非常低的核苷酸多样性（95.9–100％）。他们与从世界其他地方存放在GenBank中的马铃薯叶卷病毒分离物序列具有最高的98.08-99.36％nt同源性。系统进化分析和外壳蛋白基因序列的单倍型网络表明，塞尔维亚马铃薯卷叶病毒分离株可以分为两个不同的组，表明该病毒可能被引入了塞尔维亚。这项研究的结果证实了马铃薯卷叶病毒在塞尔维亚种薯生产中的重要性。