本文選題：豬瘟病毒　切入點(diǎn)：野毒　出處：《中國畜牧獸醫(yī)》2016年09期 　論文類型：期刊論文

【摘要】：為建立一種對(duì)豬瘟病毒(classical swine fever virus,CSFV)臨床樣本快速、簡便的檢測技術(shù),以區(qū)分豬瘟(classical swine fever,CSF)野毒和疫苗毒感染,為規(guī)�；i場CSF凈化奠定基礎(chǔ)。通過對(duì)GenBank中CSF野毒、疫苗毒及近源病毒的基因組全序列比對(duì),發(fā)現(xiàn)CSF兔化弱毒疫苗株3′-NTR獨(dú)立存在富含T的插入序列,根據(jù)這一特點(diǎn)分別在該插入序列的上、下兩端設(shè)計(jì)2對(duì)單一RT-PCR引物并選擇特異保守區(qū)域設(shè)計(jì)了二重RT-PCR引物,優(yōu)化篩選能夠鑒別CSF野毒與兔化弱毒疫苗的PCR反應(yīng)條件,建立了能鑒別CSF野毒和疫苗毒的單一與二重RT-PCR鑒別診斷方法。敏感性和特異性分析表明,單一RT-PCR檢測CSFV各引物最低核酸檢測量分別為2.2pg(單一RT-PCR中的1對(duì)引物)和1.7pg(單一RT-PCR中的另外1對(duì)引物);二重RT-PCR檢測CSFV各引物最低核酸檢測量分別為8.2pg(CSF野毒)和6.7pg(CSF疫苗毒),兩種方法均檢測不到PRV、PRRSV、JEV、BVDV、PCV2的DNA/RNA。采用該方法對(duì)146份可疑臨床樣品進(jìn)行檢測,結(jié)果表明CSF疫苗毒與野毒在能繁母豬、育肥豬、保育豬和哺乳仔豬中的二重感染率分別為6.3%、7.4%、8.3%、8.6%。本研究建立的單一與二重RT-PCR方法都具有敏感性強(qiáng)、特異性優(yōu)、重復(fù)性好的特點(diǎn),該研究對(duì)規(guī)模化豬場豬瘟的凈化具有十分重要的參考價(jià)值。
[Abstract]:In order to establish a rapid and simple method for the detection of classical swine fever virus (CSFV) from classical swine fever virus (CSFV), and to distinguish the field and vaccine infection of CSFV, and to lay a foundation for the purification of CSFV on a large scale, CSF wild virus in GenBank was obtained. The whole genome sequence alignment of vaccine virus and near-source virus showed that there was T-rich insertion sequence in CSF attenuated rabbit-attenuated vaccine strain 3ntr. According to this characteristic, the insertion sequence was found. Two pairs of single RT-PCR primers were designed at both ends of the study, and the specific conserved region was selected to design double RT-PCR primers. The conditions of PCR reaction were optimized for the identification of CSF wild virus from rabbit attenuated vaccine. A single and double RT-PCR differential diagnosis method for differentiating CSF wild virus from vaccine virus was established. The sensitivity and specificity analysis showed that, The minimum nucleic acids detected by single RT-PCR detection CSFV primers were 2.2 PG (one pair of primers in a single RT-PCR) and 1.7 PG (another pair of primers in a single RT-PCR; the lowest nucleic acid detection of each primer of double RT-PCR for CSFV was 8.2 PG CSF wild virus) and 6.7 pgCSF were detected respectively. The DNA / RNA of PRVV VV VV VV2 PCV2 was not detected by both methods. 146 suspicious clinical samples were detected by this method. The results showed that the double infection rates of CSF vaccine and wild virus in fertile sows, fattening pigs, care pigs and suckling piglets were 6. 3% and 7. 4% and 8. 6%, respectively. The single and double RT-PCR methods developed in this study were highly sensitive, specific and reproducible. This study has very important reference value for the purification of swine fever in scale pig farm.
【作者單位】： 貴州大學(xué)動(dòng)物科學(xué)學(xué)院;貴州省動(dòng)物疫病預(yù)防控制中心;
【基金】：貴州省2014年農(nóng)業(yè)攻關(guān)項(xiàng)目(黔科合NY字[2014]3055號(hào)) 貴州省科學(xué)技術(shù)基金項(xiàng)目(黔科合J字[2013]2110號(hào)) 貴州大學(xué)研究生創(chuàng)新基金(農(nóng)研[2015]030號(hào))
【分類號(hào)】：S852.651

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