Department of Laboratory Animal Center, Liaoning Medical University, Jinzhou, China.
PLoS One. 2013;8(2):e52670. doi: 10.1371/journal.pone.0052670. Epub 2013 Feb 8.
Sacbrood virus (SBV) is a picorna-like virus that affects honey bees (Apis mellifera) and results in the death of the larvae. Several procedures are available to detect Chinese SBV (CSBV) in clinical samples, but not to estimate the level of CSBV infection. The aim of this study was develop an assay for rapid detection and quantification of this virus. Primers and probes were designed that were specific for CSBV structural protein genes. A TaqMan minor groove binder (MGB) probe-based, fluorescence real-time quantitative PCR was established. The specificity, sensitivity and stability of the assay were assessed; specificity was high and there were no cross-reactivity with healthy larvae or other bee viruses. The assay was applied to detect CSBV in 37 clinical samples and its efficiency was compared with clinical diagnosis, electron microscopy observation, and conventional RT-PCR. The TaqMan MGB-based probe fluorescence real-time quantitative PCR for CSBV was more sensitive than other methods tested. This assay was a reliable, fast, and sensitive method that was used successfully to detect CSBV in clinical samples. The technology can provide a useful tool for rapid detection of CSBV. This study has established a useful protocol for CSBV testing, epidemiological investigation, and development of animal models.
Sacbrood 病毒（SBV）是一种类似微小 RNA 病毒的病毒，会导致幼虫死亡。有几种方法可用于检测临床样本中的中国 Sacbrood 病毒（CSBV），但无法估计 CSBV 感染的程度。本研究旨在开发一种用于快速检测和定量分析该病毒的方法。设计了针对 CSBV 结构蛋白基因的引物和探针。建立了基于 TaqMan 小沟结合物（MGB）探针的荧光实时定量 PCR。评估了该方法的特异性、灵敏度和稳定性；特异性高，与健康幼虫或其他蜜蜂病毒无交叉反应。该方法应用于检测 37 个临床样本中的 CSBV，并将其效率与临床诊断、电子显微镜观察和常规 RT-PCR 进行了比较。TaqMan MGB 探针荧光实时定量 PCR 法比其他测试方法更灵敏。该方法是一种可靠、快速、灵敏的方法，成功地用于检测临床样本中的 CSBV。该技术为快速检测 CSBV 提供了有用的工具。本研究为 CSBV 的检测、流行病学调查和动物模型的建立建立了一个有用的方案。
