【摘要】 为了建立一种可同时检测区分猪瘟病毒(CSFV),猪繁殖与呼吸障碍综合征病毒(PRRSV)和猪圆环病毒2型(PCV-2)的寡核苷酸基因芯片方法,本研究针对每种病毒设计了4条～6条寡核苷酸探针,检测低限为1.6×104PCV-2拷贝数/μL,1.6×104CSFV拷贝数/μL,1.6×105PRRSV拷贝数/μL,比琼脂糖凝胶灵敏约10倍。利用建立的寡核苷酸基因芯片方法对76个仔猪样本进行了检测,检出了3种病毒的存在,其中25个样本(32.9%)同时感染了2种以上病毒。结果表明寡核苷酸基因芯片检测是一种快速、灵敏和高效的猪只混合感染病毒的病原学诊断方法。更多还原

【Abstract】 Porcine circovirus type 2 (PCV-2),classical swine fever virus (CSFV) and porcine reproductive and respiratory syn-drome virus (PRRSV) are important viruses that cause severe reproductive and/or respiratory failure in swine,and rapid and accu-rate detection and identification of these pathogens is critical for swine disease management. In the present study,we developed a multiplex polymerase chain reaction (PCR) based oligonucleotide microarray assay that included four to six specific oligonu-cleotides for each virus. The microarray was tested on three target viruses and 76 clinical specimens. The detection limit of the oligonucleotide microarray was 1.6×104,1.6×104,1.6×105 copies/μL for PCV-2,CSFV and PRRSV,respectively,which was 10-fold more sensitive than that of agarose gel. Co-infection with two or three viruses was detected in 26 specimens (32.8 %). Theresults indicated that the microarray detection system is a reliable and effective method for viral pathogen detection and identification even when multiple pathogens are present in a porcine clinical sample.更多还原