安卡拉病毒PCR检测方法的建立

Establishment and Application of PCR Method for Ankara Virus

由安卡拉病毒引起的鸡心包积液-肝炎综合征发病急,传播速度快,造成雏鸡的高死亡率。为快速确诊,实现早发现早控制,本试验根据安卡拉病毒主要结构蛋白六邻体(Hexon)基因序列,设计1对特异性引物,优化反应体系,并对其进行特异性、敏感性和临床样本检测,以期建立检测安卡拉病毒的PCR方法。结果显示,可扩增出295 bp特异性片段,最佳模板质量浓度确定为4.38×10^(-2) g/L,最佳退火温度为58℃。该方法具有较好的特异性、灵敏性和临床应用性,可对临床FAdV-4感染病例作出快速检测,为疫病的快速鉴别诊断、及时防控提供参考价值。

The chicken heart effusion-hepatitis syndrome caused by the Ankara virus is acute and spreads quickly,resulting in a high mortality rate for chicks.To achieve rapid diagnosis and early detection and control,we designed a pair of specific primers based on the sequence of the main structural protein Hexon of Ankara virus,optimized the reaction system,and tested its specificity,sensitivity and clinical samples to establish a PCR method for detection of Ankara virus.The results showed that 295 bp specific fragment was amplified.The optimal template mass concentration was determined to be 4.38×10^(-2) g/L,and the optimal annealing temperature was 58℃.This method has good specificity,sensitivity and clinical application,and can be used for rapid detection of FAdV-4 infection in clinical cases.