CAJ | 학술논문

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 × 10~4 PCV-2拷贝数/μL,1.6 × 10~4 CSFV拷贝数/μL,1.6×10~5 PRRSV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.
위료건립일충가동시검측구분저온병독(CSFV),저번식여호흡장애종합정병독(PRRSV)화저원배병독2형(PCV-2)적과핵감산기인심편방법,본연구침대매충병독설계료4조～6조과핵감산탐침,검측저한위1.6 × 10~4 PCV-2고패수/μL,1.6 × 10~4 CSFV고패수/μL,1.6×10~5 PRRSV고패수/μL,비경지당응효령민약10배.이용건립적과핵감산기인심편방법대76개자저양본진행료검측,검출료3충병독적존재,기중25개양본(32.9%)동시감염료2충이상병독.결과표명과핵감산기인심편검측시일충쾌속、령민화고효적저지혼합감염병독적병원학진단방법.
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 × 10~4,1.6 × 10~4,1.6×10~5 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%).The results indicated that the microarray detection system is a reliable and effective method for viral pathogen detection and identifica-tion even when multiple pathogens are present in aporcine clinical sample.