用改良的RT-PCR技术快速检测传染性法氏囊病病毒

Rapid detection of infectious bursal disease virus by a novel reverse transcription polymerase chain reaction amplification assay

本研究将最新的病毒核酸纯化技术和单管ＲＴ－ＰＣＲ技术相结合，应用于ＩＢＤＶ的诊断研究，对４０余个病毒样品进行扩增反应，均取得令人满意的效果。该技术灵敏快速，从核酸纯化到电脉检测ＰＣＲ产物只需５～６ｈ，反应灵敏度比传统方法至少提高１００倍。所设计的引物对毒株的适用范围较广。

A novel reverse transcription polymerase chain reaction (RTPCR)amplification assay was developed to detect infectious bursal disease virus (IBDV) gene sequences in bursal samples and cell cultures.One pair of primer were designed to amplify a fragment of segment A gene that partialy code for the IBDV protiens VP4.Analysis of 12 strains of IBDV and 12 bursal samples all yielded the specific 365bp fragments.The assay was a combination of a new developed virus RNA purification technique and TitanTm one tube RTPCR system,that could gain detecting result within 5￣6 hours.It was 100 times more sensitive than the tranditional two step two tube RTPCR assay,and was easy to perform thus is suitable for the treatment of large samples in animal quarantine.