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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.

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Twenty-five gilts without measurable serum antibody titres to porcine reproductive and respiratory syndrome virus (PRRSV) were identified and 16 were inoculated with PRRSV at seven, 14 or 21 days of gestation and killed 20 to 22 days later to determine the effect of the virus on their embryos. The remaining nine gilts were not exposed to PRRSV, but were killed at the same stages of gestation. The gilts were observed for clinical signs of infection and the gilts and their embryos were tested for PRRSV and homologous antibodies. The infection was demonstrated by the re-isolation of the virus and its detection by the reverse transcriptase polymerase chain reaction in serum and other tissue samples from the inoculated gilts, and also by seroconversion. However, the gilts remained healthy throughout the study, except for one which was depressed and anorexic for two days. Two of the litters from the gilts challenged with PRRSV on day 14 of gestation contained one and three infected live embryos; the other embryos from these two litters did not contain detectable virus, although most of the embryos in one of the litters were dead. The other nine litters from the gilts challenged with PRRSV and the control litters, showed no evidence of infection.

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