PDF(Pubmed)
Abstract：
UNASSIGNED: As an important social insect, honey bees play crucial roles in agricultural production, sustainable development of agricultural production, and the balance of the natural environment. However, in recent years, Israeli acute paralysis virus (IAPV) and chronic bee paralysis virus (CBPV), the main pathogens of bee paralysis, have continuously harmed bee colonies and caused certain losses to the beekeeping industry. Some beekeeping farms are located in wild or remote mountainous areas, and samples from these farms cannot be sent to the laboratory for testing in a timely manner, thereby limiting the accurate and rapid diagnosis of the disease.
UNASSIGNED: In this study, we used a reverse transcription-recombinase polymerase amplification-lateral flow dipstick (RT-RPA-LFD) method for the dual detection of IAPV and CBPV. RPA primers and LFD detection probes were designed separately for their conserved genes. Primers and probes were screened, and the forward and reverse primer ratios, reaction times, and temperatures were optimized. According to the results of the optimization tests, the optimal reaction temperature for RT-RPA is 37°C, and when combined with LFD, detection with the naked eye requires <20 min. The developed RPA-LFD method specifically targets IAPV and CBPV and has no cross-reactivity with other common bee viruses. In addition, the minimum detection limit of the RT-RPA-LFD method is 101 copies/μL.
UNASSIGNED: Based this study, this method is suitable for the detection of clinical samples and can be used for field detection of IAPV and CBPV.
UNASSIGNED: In this study, we used a reverse transcription-recombinase polymerase amplification-lateral flow dipstick (RT-RPA-LFD) method for the dual detection of IAPV and CBPV. RPA primers and LFD detection probes were designed separately for their conserved genes. Primers and probes were screened, and the forward and reverse primer ratios, reaction times, and temperatures were optimized. According to the results of the optimization tests, the optimal reaction temperature for RT-RPA is 37°C, and when combined with LFD, detection with the naked eye requires <20 min. The developed RPA-LFD method specifically targets IAPV and CBPV and has no cross-reactivity with other common bee viruses. In addition, the minimum detection limit of the RT-RPA-LFD method is 101 copies/μL.
UNASSIGNED: Based this study, this method is suitable for the detection of clinical samples and can be used for field detection of IAPV and CBPV.
摘要：
■作为一种重要的社会性昆虫,蜜蜂在农业生产中起着至关重要的作用，农业生产的可持续发展,和自然环境的平衡。然而,近年来,以色列急性麻痹病毒（IAPV）和慢性蜜蜂麻痹病毒（CBPV），蜜蜂麻痹的主要病原体，不断危害蜂群，给养蜂业造成一定损失。一些养蜂场位于野生或偏远山区,这些农场的样品不能及时送到实验室进行检测，从而限制了疾病的准确和快速诊断。
■在这项研究中，我们使用逆转录-重组酶聚合酶扩增-侧流试纸(RT-RPA-LFD)方法双重检测IAPV和CBPV.针对其保守基因分别设计了RPA引物和LFD检测探针。引物和探针进行筛选，以及正向和反向引物比率，反应时间,和温度进行了优化。根据优化试验的结果,RT-RPA的最佳反应温度为37°C，当与LFD结合时，用肉眼检测需要<20分钟。开发的RPA-LFD方法专门针对IAPV和CBPV，与其他常见的蜜蜂病毒没有交叉反应性。此外,RT-RPA-LFD方法的最低检测限为101拷贝/μL。
■基于这项研究，该方法适用于临床样品的检测，可用于IAPV和CBPV的现场检测。
■在这项研究中，我们使用逆转录-重组酶聚合酶扩增-侧流试纸(RT-RPA-LFD)方法双重检测IAPV和CBPV.针对其保守基因分别设计了RPA引物和LFD检测探针。引物和探针进行筛选，以及正向和反向引物比率，反应时间,和温度进行了优化。根据优化试验的结果,RT-RPA的最佳反应温度为37°C，当与LFD结合时，用肉眼检测需要<20分钟。开发的RPA-LFD方法专门针对IAPV和CBPV，与其他常见的蜜蜂病毒没有交叉反应性。此外,RT-RPA-LFD方法的最低检测限为101拷贝/μL。
■基于这项研究，该方法适用于临床样品的检测，可用于IAPV和CBPV的现场检测。
