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目的建立可同时检测新城疫病毒(Newcastle disease virus,NDV)和传染性支气管炎病毒(Infectious bronchitis virus,IBV)的多重DNA微列阵鉴定方法。方法针对NDV和IBV设计PCR引物和微阵列探针,制备检测用微阵列。将样品与微列阵进行杂交,分析微阵列的特异性和灵敏度。结果建立的微阵列检测NDV和IBV为阳性,且能区分NDV和IBV,而流感病毒传染性法氏囊病毒等阴性对照无杂交信号。通过GenePix 4.1软件分析杂交信号的平均SNR532,NDV和IBV的检出限为1×10~3copies/μl。结论建立的多重DNA微列阵鉴定方法具有高特异性和高敏感性,可用于NDV和IBV感染检测。
Objective To establish a multiplex DNA microarray assay for the simultaneous detection of Newcastle disease virus (NDV) and infectious bronchitis virus (IBV). Methods PCR primers and microarray probes were designed for NDV and IBV to prepare detection microarrays. The sample is hybridized to the microarray to analyze the specificity and sensitivity of the microarray. Results The microarray was positive for NDV and IBV, and NDV and IBV were distinguishable. However, no negative signal was detected in the negative control such as infectious bursal disease virus. The average SNR532 of hybridization signals was analyzed by GenePix 4.1 software. The detection limits of NDV and IBV were 1 × 10 ~ 3 copies / μl. Conclusion The established multiplex DNA microarray identification method has high specificity and high sensitivity and can be used for the detection of NDV and IBV infection.