CAJ | 학술논문

为建立一种具有敏感性高、特异性强、方便、快速等特点的检测猪繁殖与呼吸综合征病毒（PRRSV ）的环介导等温扩增技术（LAM P），针对美洲型PRRSV 的ORF5基因设计了一套LAM P引物，建立了LAM P反应体系，并对其敏感性、特异性等进行了评估。结果表明，该方法可在65℃、1 h内完成最低限度5．49 pg/μL含量的PRRSV核酸扩增，结果仅肉眼就能判定。该检测方法特异性强，与猪瘟病毒（CS-FV ）、猪圆环病毒2型（PCV-2）和猪伪狂犬病病毒（PRV ）等病原体无交叉反应；对25份临床样品检测表明，该方法的检出率高于常规RT-PCR法，且操作更为方便、快速。该检测方法特异、敏感、准确，可应用于临床检测PRRSV。
위건립일충구유민감성고、특이성강、방편、쾌속등특점적검측저번식여호흡종합정병독（PRRSV ）적배개도등온확증기술（LAM P），침대미주형PRRSV 적ORF5기인설계료일투LAM P인물，건립료LAM P반응체계，병대기민감성、특이성등진행료평고。결과표명，해방법가재65℃、1 h내완성최저한도5．49 pg/μL함량적PRRSV핵산확증，결과부육안취능판정。해검측방법특이성강，여저온병독（CS-FV ）、저원배병독2형（PCV-2）화저위광견병병독（PRV ）등병원체무교차반응；대25빈림상양품검측표명，해방법적검출솔고우상규RT-PCR법，차조작경위방편、쾌속。해검측방법특이、민감、준학，가응용우림상검측PRRSV。
To develop a loop-mediated isothermal amplification method (LAM P ) with high sensitivity , good specificity ,convenience and fastness for detection of porcine reproductive and respiratory syndrome virus (PRRSV) ,a set of LAMP primers were designed according to the ORF5 gene of PRRSV American type and a LAMP reaction system was established .The sensitivity and specificity of the system were eval-uated .The results showed that the assay could carry out nucleic acid amplification reaction of PRRSV within 1 h under 65℃ with the detection limit of 5 .49 pg/μL and the results could be read by naked eye without gel electrophoresis .This detection method was highly specific ,and had no cross reaction with pathogens such as classical swine fever virus (CSFV) ,porcine circovirus type 2 (PCV-2) and porcine pseu-dorabies virus(PRV);detection results of 25 clinical samples showed that the detection rate of this assay was higher than that with conventional RT-PCR and this assay operated quickly and conveniently .This de-tection method was specific ,sensitive and accurate ,and could be used for clinical detection of PRRSV .