A real time RT-PCR assay for the specific detection of Peste des petits ruminants virus.
Peste des petits ruminants virus (PPRV) causes a devastating disease of small ruminants present across much of Africa and Asia. Recent surveillance activities and phylogenetic analyses have suggested that the virus is an emerging problem as it is now being detected in areas previously free of the disease. As such, the virus not only is threatening small ruminant production and agricultural stability in the developing world, but also poses an economic threat to livestock in the European Union (EU) through introduction from European Turkey and North Africa. This report describes the development of a high throughput, rapid, real time RT-PCR method for the sensitive and specific detection of PPRV using robotic RNA extraction. This assay targets the nucleocapsid (N) gene of PPRV and has been shown to detect all four genetic lineages of PPRV in tissues, ocular and nasal swabs and blood samples collected in the field. The lowest detection limit achieved was approximately 10 genome copies/reaction, making this assay an ideal tool for the sensitive and rapid detection of PPRV in diagnostic laboratories.
Institute for Animal Health, Ash Road, Pirbright, Woking, Surrey GU240NF, UK. carrie.batten@bbsrc.ac.uk
Carrie A
CA
Batten
Ashley C
AC
Banyard
Donald P
DP
King
Mark R
MR
Henstock
Lorraine
L
Edwards
Anna
A
Sanders
Hubert
H
Buczkowski
Chris C L
CC
Oura
Tom
T
Barrett
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D019251:Nucleocapsid / Q000235:genetics; D029021:Peste-des-Petits-Ruminants / Q000175:diagnosis* / Q000821:virology; D018107:Peste-des-petits-ruminants virus / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012418:Ruminants / Q000821:virology*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
D017931:DNA Primers