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以蓝莓叶片为试材,提取病毒RNA,根据已报道的蓝莓休克病毒(Blueberry shock virus,BlShV)外壳蛋白基因序列设计特异性引物,建立RT-PCR快速检测方法,并对采集的20份疑似病例进行检测。结果表明:建立的RT-PCR方法具有良好的特异性和较高的灵敏度。仅从感染BlShV的样品中扩增出与预期大小相符的特异性目的片段,而从其它蓝莓病毒及健康样品中未扩增到目的片段;该方法能检测到RNA原液的10-4倍稀释液,灵敏度较高。疑似病例检测结果与ELISA验证结果完全相符。试验表明,建立的RT-PCR方法能够用于BlShV的现地检测。
Using blueberry leaves as test material, the viral RNA was extracted, and specific primers were designed according to reported blueberry shock virus (BlShV) coat protein gene sequences. A rapid RT-PCR method was established and 20 suspected cases Test. The results showed that the established RT-PCR method has good specificity and high sensitivity. Only the specific fragment of interest that matches the expected size was amplified from the BlShV-infected sample, but not amplified from other blueberries and healthy samples. The method can detect 10-4-fold dilution of RNA stock solution , High sensitivity. Suspected case test results and ELISA validation results exactly. Experiments show that the established RT-PCR method can be used for the in situ detection of BlShV.