為探索適合臨床高通量雞傳染性喉氣管炎檢測的方法,針對雞傳染性喉氣管炎病毒gB基因序列保守區(qū)域,,篩選1對特異性引物和1條特異性探針,建立了實(shí)時熒光PCR檢測方法,,并對該方法進(jìn)行特異性,、敏感性評估和臨床應(yīng)用檢測。結(jié)果顯示:篩選出的引物和探針與其他禽類常見病毒無交叉反應(yīng),,檢測下限達(dá)到100拷貝/反應(yīng),;對來自13個場的480份臨床樣本進(jìn)行檢測,檢測出17個陽性樣品,,與常規(guī)PCR檢測結(jié)果符合率為100%,。結(jié)果表明,此方法特異性強(qiáng),,靈敏度高,、耗時少,可用于雞傳染性喉氣管炎的快速檢測,。
Establishment and Application of a Real-time PCR for Detecting Infectious Laryngotracheitis Virus
In order to develop an appropriate method for high-throughput detection of avian infectious laryngotracheitis(AILT),,a real-time PCR assay was established with a pair of specific primers and a probe based on the conservative zone of gB gene sequence of AILT,then its specificity and sensitivity were evaluated,,and its clinical application was tested. The results showed that the selected primers and probe failed to cross-react with other common poultry viruses,,with the detection limit of 100 copies/reaction. 17 out of 480 clinical samples collected from 13 farms were positive. The results completely conformed to the test result by common PCR assay. As a conclusion,AILT could be rapidly detected by the established method with high specificity,,high sensitivity and less time-consuming.
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