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豬德爾塔冠狀病毒納米PCR檢測方法的建立

點擊數:6638  時間:2019-08-13 16:56:40  來源: 信息處      

根據GenBank登錄的豬德爾塔冠狀病毒(PDCoV)毒株M基因設計特異性引物,建立了PDCoV納米PCR檢測方法并進行了臨床樣品檢測。結果顯示:該方法對标準品檢測的最低濃度可達102 copies/μL,與其他幾種常見豬病病毒無交叉反應;用該方法檢測國内豬場62份腹瀉糞便樣品,相比常規PCR方法,該納米PCR方法的檢出率更高,而兩種方法對268份進境活豬糞拭子樣品檢測均為陰性。結果表明,本研究所建立的納米PCR檢測方法适用于國内豬場的PDCoV檢測和進境活豬的監控檢測,可作為PDCoV實驗室檢測的有力技術手段。

Establishment of Nanoparticle-assisted PCR Assay for Detection ofPorcine Deltacoronavirus

In this paper,a pair ofprimers were designed according to the M gene sequence of porcinedeltacoronavirus(PDCoV)that waspublished in Genbank,then a kind ofnanoparticle-assisted PCR assay was established. The results showed that thestandard samples with minimum concentration of 102 copies/μL could be detected by this method,and therewas no any cross-reaction with other common swine disease viruses. Compared toregular PCR assay,the nanoparticle-assisted PCR assaypresented a higher detection rate when 62 diarrhea feces collected fromdomestic swine farms were tested. When 268 fecal swab samples of imported livepigs were tested by both the two methods,the detectionresults all showed negative. Therefore,thenanoparticle-assisted PCR assay established in the paper could be used indetecting PDCoV in domestic swine farms and monitoring imported live pigs,which would be a strong technical tool for laboratory detection ofPDCoV.

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