Nested PCR for detection and typing of porcine reproductive and respiratory syndrome (PRRS) virus in pigs.
A nested polymerase chain reaction (PCR) was developed to detect porcine reproductive and respiratory syndrome (PRRS) virus. A common primer set for European and North American type isolates of PRRS virus was designed for reverse transcription PCR, and a specific primer set for each of the 2 type isolates was designed for nested PCR. The PCR that used a specific primer set detected the corresponding type of the virus at a level equivalent to 1 TCID50/100 microliters, but not the other type of isolates. Therefore, the method clearly differentiated the 2 types of virus from each other. The detection of PRRS virus by the nested PCR was as sensitive as virus isolation in cultures of porcine alveolar macrophages from infected pigs at the acute stages, and was more sensitive from pigs at the convalescent stages. The infecting virus type was determined by use of 2 specific primer sets even when virus isolation was negative in naturally infected pigs. It was concluded that the nested PCR is useful for diagnosis and typing of PRRS virus and studies of persistent infection by the virus.
Exotic Diseases Research Division, National Institute of Animal Health, Tokyo, Japan.
Y
Y
Kono
T
T
Kanno
M
M
Shimizu
S
S
Yamada
S
S
Ohashi
M
M
Nakamine
J
J
Shirai
D000818:Animals; D017931:DNA Primers; D016133:Polymerase Chain Reaction; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis*; D019316:Porcine respiratory and reproductive syndrome virus / Q000145:classification* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013552:Swine / Q000821:virology
D017931:DNA Primers