Bornova/Izmir Veterinary Control Institute, Department of Virology, Bornova, Izmir, 35040, Turkey.
Ondokuz Mayis University, Faculty of Veterinary Medicine, Department of Virology, Samsun, 55139, Turkey.
J Virol Methods. 2020 Jul;281:113858. doi: 10.1016/j.jviromet.2020.113858. Epub 2020 Mar 20.
Colony losses in apiaries are frequently one of the most important problems in beekeeping. Colony loss is multifactorial with many reported disorders, Colony Collapse Disorder (CCD), is an increasingly recognised phenomenon which is thought to be caused by many pathogens, including viruses. The aim of this study was to develop a multiplex RT-PCR (mRT-PCR) test to obtain faster results in routine diagnostic laboratories for seven crucial bee viruses. Specific primers for seven RNA viruses, including Israeli acute bee paralysis virus (IAPV), deformed wing virus (DWV), sacbrood virus (SBV), acute bee paralysis virus (ABPV), black queen cell virus (BQCV), kashmir bee virus (KBV) and chronic paralysis virus (CBPV), were used for testing procedure. The mRT-PCR assay can amplify seven plasmid DNA fragments from the pooled viral genomes and it was shown to be sensitive because virus copy numbers were detected to be 10 copies/μl when log serial dilutions were performed for the optimized mRT- PCR method. It is concluded that, mRT-PCR test can be used in routine analysis because this assay can perform specific, sensitive and reliable results also achieves economic gains and time due to detecting seven viral agents simultaneously.
蜂群损失是养蜂业中最常见的问题之一。蜂群损失是多因素的，有许多报道的疾病，包括病毒性疾病。本研究旨在开发一种多重 RT-PCR（mRT-PCR）检测方法，以便在常规诊断实验室中更快地获得七种关键蜜蜂病毒的检测结果。本研究使用了针对七种 RNA 病毒的特异性引物，包括以色列急性麻痹病毒（IAPV）、变形病毒（DWV）、囊状幼虫病病毒（SBV）、急性麻痹病毒（ABPV）、黑皇后细胞病毒（BQCV）、克什米尔蜜蜂病毒（KBV）和慢性麻痹病毒（CBPV）。mRT-PCR 检测方法可以从混合的病毒基因组中扩增出七个质粒 DNA 片段，并且具有较高的敏感性，因为当对优化的 mRT-PCR 方法进行对数系列稀释时，可以检测到 10 个拷贝/μl 的病毒拷贝数。研究结果表明，mRT-PCR 检测方法可用于常规分析，因为该方法可以同时检测七种病毒，具有特异性、敏感性和可靠性，并且可以节省经济成本和时间。
