【摘要】：我國(guó)當(dāng)前豬流行性腹瀉病毒(PEDV)流行毒株與經(jīng)典毒株相比,在S基因上存在多處變異,其中在58aa處有4個(gè)氨基酸QGVN的插入。為快速鑒別經(jīng)典毒株與變異毒株,本試驗(yàn)根據(jù)Gen Bank中當(dāng)前流行毒株S基因序列設(shè)計(jì)合成2對(duì)特異性擴(kuò)增引物,在優(yōu)化RT-PCR反應(yīng)條件的基礎(chǔ)上,建立一套區(qū)分PEDV變異毒株與經(jīng)典毒株的套式RT-PCR檢測(cè)方法,并完成特異性、敏感性試驗(yàn)及對(duì)臨床送檢樣品檢測(cè)試驗(yàn)。結(jié)果表明:該套式PCR檢測(cè)方法可以分別鑒定PEDV經(jīng)典毒株和變異毒株,PEDV經(jīng)典毒株僅在PCR外套產(chǎn)物有749 bp的條帶,內(nèi)套產(chǎn)物沒有條帶;而PEDV變異毒株在PCR外套產(chǎn)物中有760 bp的條帶,而在內(nèi)套產(chǎn)物中出現(xiàn)510 bp的條帶。該檢測(cè)方法特異性強(qiáng)、靈敏度高、操作簡(jiǎn)單,為豬流行性腹瀉的快速診斷提供了一種較為方便的技術(shù)手段。
[Abstract]:Compared with the classical strains of porcine epidemic diarrhea virus (PEDV) in China, there are many mutations in S gene, among which four amino acids QGVN are inserted at 58aa. In order to quickly identify the classical and variant strains, two pairs of specific primers were designed and synthesized according to the S gene sequence of the current prevalent strain in GenBank. Based on the optimization of the conditions of RT-PCR reaction, two pairs of primers were designed and synthesized. A nested RT-PCR method was established to distinguish PEDV variant strains from classical strains, and the specificity, sensitivity and clinical samples were tested. The results showed that the classical PEDV strain and the mutant strain PEDV could be identified by nested PCR, only 749 BP bands were found in the PCR coat product, and there were 760 BP bands in the PCR coat product of PEDV mutant strain, and there were 749 BP bands in the PCR coat product, and 760 BP band in the PCR coat product of the PEDV mutant strain. A 510 BP band was found in the inner product. This method has the advantages of high specificity, high sensitivity and simple operation, which provides a more convenient technique for the rapid diagnosis of swine epidemic diarrhea.
【作者單位】： 華南農(nóng)業(yè)大學(xué)獸醫(yī)學(xué)院;廣東省動(dòng)物源性人獸共患病預(yù)防與控制重點(diǎn)實(shí)驗(yàn)室;
【基金】：廣東省生豬產(chǎn)業(yè)體系創(chuàng)新團(tuán)隊(duì)項(xiàng)目
【分類號(hào)】：S852.651

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