梨火疫细菌实时荧光PCR和诱捕PCR-ELISA检测方法的建立

The establishment of detection of real-time fluorescent PCR and capture-PCR-ELISA for detection of fire blight Erwinia amylovora.

根据梨火疫细菌中独特而保守存在的质粒pEA2 9,设计了 1对引物和 3条探针 ,建立了实时荧光PCR检测方法和诱捕PCR ELISA检测方法。实时荧光PCR采用带荧光标记的核酸杂交探针 ,边扩增边检测 ,步骤简单 ,不需PCR后处理 ,可避免假阳性和交叉污染 ;诱捕PCR ELISA检测方法只需简单处理的样品就能检测 ,减少了核酸不纯出现的漏检 ,由于增加了核酸杂交探针 ,可不需凝胶电泳EB染色检测 ,不会出现假阳性问题。

A pair of primer and three probes on the nucleotide sequence of pasmid pEA29 which is known to be unique to and conserved in Erwinia amylovora,the novel real time fluorescent PCR and capture PCR ELISA were established to detect E amylovora The real time fluorescent PCR use a fluorescent probe,detection and PCR amplification are the same time,which has many advantages:the simple step,no post PCR manipulations and reducing false positive result and contamination risks The capture PCR ELISA only need simply sample handled and can reduce non detectable results Due to hybridization probe eliminating ethidium bromide staining and gel running also reducing false positive