돼지 호흡기 생식기증 바이러스(porcine reproductive and respiratory syndrome virus: PRRSV) 감염이 의심되는 농장으로부터 수집된 혈청가검물 646개로부터 MARC-145 cell을 이용하여 PRRSV 분리를 시도한 바 MARC-145 세포단층상에 세포변성효과(cytopathic effects : CPE)를 나타내는 바이러스 36주를 분리하였다. 분리된 36주가 PRRSV인지 여부를 확인하기 위하여 PRRSV를 실험적으로 접종한 혈청을 이용하여 간접형광항체시험과 혈청중화시험을 실시한 결과 36주 모두가 PRRSV로 동정되었다. 혈청학적인 동정법과 더불어 reverse-transcription polymerase chain reaction을 이용하여 PRRSV open reading frame 5(ORF5)의 유전자를 증폭한 결과 선발된 6주 모두에서 80bp의 flanking sequencing를 포함하여 약 680bp의 ORF5의 유전자를 증폭할 수 있었다.
Isolation of PRRSV was attempted from 646 swine sera collected from swine farms. The MARC-145 cell, which is highly permissive to PRRSV, was used for virus isolation, propagation, IFA test, and VN test. Total 36 cytopathic viruses to MARC-145 cells were isolated. The virus isolates were identified as a PRRSV by the IFA test and VN test using the reference sera prepared by experimental infection of reference PRRSV CNV-1 into 30 day-old pig. In addition to serological conformation, ORF5 of genomic RNA of 6 selected cytopathic viruses were amplified by the RT-PCR. The resulting PCR products were examined by electrophoresis in 1.2% agarose gel. An appropriate bands of about 680bp including the flanking sequence of total 80bp were seen on agarose gel.
