Simple and rapid detection of the porcine reproductive and respiratory syndrome virus from pig whole blood using filter paper.
The combination of Flinders Technology Associates filter papers (FTA cards) and real-time PCR was examined to establish a simple and rapid technique for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) from whole pig blood. A modified live PRRS vaccine was diluted with either sterilised saline or pig whole blood, and the suspensions were applied onto the FTA cards. The real-time RT-PCR detection of PRRSV was performed directly with the samples applied to the FTA card without the RNA extraction step. Six whole blood samples from at random selected piglets in the PRRSV infected farm were also assayed in this study. The expected PCR product was successfully amplified from either saline diluted or pig whole blood diluted vaccine. The same PCR ampliocon was detected from all blood samples assayed in this study. This study suggested that the combination of an FTA card and real-time PCR is a rapid and easy technique for the detection of PRRSV. This technique can remarkably shorten the time required for PRRSV detection from whole blood and makes the procedure much easier.
Inoue
Ryo
R
Kyoto Prefectural University, Laboratory of Animal Science, 1-5 Shimogamo, Kyoto 606-8522, Japan.
Tsukahara
Takamitsu
T
Sunaba
Chinatsu
C
Itoh
Mitsugi
M
Ushida
Kazunari
K
D000818:Animals; D001483:Base Sequence; D005374:Filtration / Q000662:veterinary; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary; D010209:Paper; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis*; D012689:Sequence Homology, Nucleic Acid; D034421:Sus scrofa / Q000097:blood* / Q000821:virology*; D013997:Time Factors
D012367:RNA, Viral