Suárez P, Zardoya R, Prieto C, Solana A, Tabarés E, Bautista J M, Castro J M
Departamento de Patología Animal I, Universidad Complutense de Madrid, Facultad de Veterinaria, Spain.
Arch Virol. 1994;135(1-2):89-99. doi: 10.1007/BF01309767.
A method for direct detection of the porcine reproductive and respiratory syndrome (PRRS) virus was developed, based on reverse transcription of the viral RNA coupled to DNA amplification by polymerase chain reaction. A set of primers was designed from Lelystad virus sequence within ORF 7 encoding nucleocapsid protein. From seven Spanish field isolated strains the 312 bp amplified fragment was cloned and sequenced. Alignment with Lelystad virus sequence revealed a 96-97% homology. A maximum sensitivity of 6.7 TCID50 was achieved with the reported procedure in experimentally infected swine alveolar macrophages cultures. The sensitivity obtained in crude clinical samples from experimentally infected 3-weeks old pigs was approximately 10(2) TCID50. High specificity for the PRRS virus was demonstrated for the method, as none of the seven common swine virus assayed rendered DNA amplification product.
开发了一种直接检测猪繁殖与呼吸综合征（PRRS）病毒的方法，该方法基于病毒RNA的逆转录并通过聚合酶链反应进行DNA扩增。根据编码核衣壳蛋白的开放阅读框7内的莱利斯塔德病毒序列设计了一组引物。从七个西班牙田间分离株中克隆并测序了312 bp的扩增片段。与莱利斯塔德病毒序列比对显示同源性为96-97%。在所报道的方法中，在实验感染的猪肺泡巨噬细胞培养物中实现了6.7 TCID50的最大灵敏度。在来自实验感染的3周龄仔猪的粗临床样本中获得的灵敏度约为10(2) TCID50。该方法对PRRS病毒具有高度特异性，因为所检测的七种常见猪病毒均未产生DNA扩增产物。
