Simultaneous Detection and Identification of Peste des petits ruminants
Virus Types Ⅱ and Ⅳ by MCA-Based Real-Time RT-qPCR Assay within Single
Reaction
Abstract
Peste des petits ruminants (PPR) disease is a cross-species infectious
disease that severely affects small ruminants and causes great losses to
livestock industries in various countries. Distinguishing
vaccine-immunized animals from naturally infected animals is an
important prerequisite for the eradication of PPR.
[ ](javascript:void(0);)Although peste des petits ruminants virus
(PPRV) was currently divided into lineages I-IV and only one vaccine
strain N75 belongs to type II, its epidemic strains in China all belong
to lineage IV. As to achieve this goal, we developed an SYBR Green I
real-time RT-qPCR method for rapid detection and differentiation of PPRV
lineage II and IV by analyzing different melting curve analyses. Primers
targeting the L gene were highly specific, as evidenced by the negative
amplification of closely related viruses, such as Orf virus, goat
poxvirus, and Foot-and-mouth disease virus. The sensitivity of the assay
was assessed based on plasmid DNA and the detection limit achieved was
10 copies of PPRV-II and PPRV-IV respectively. As the method has high
sensitivity, specificity, and reproducibility, which will be significant
in effectively controlling the occurrence and transmission of PPRV.