Arch Virol (2004) 149: 2155–2170
A sandwich-ELISA for the diagnosis of Peste des petits
ruminants (PPR) infection in small ruminants using
anti-nucleocapsid protein monoclonal antibody
R. P. Singh, B. P. Sreenivasa, P. Dhar, and S. K. Bandyopadhyay
Division of Virology, Indian Veterinary Research Institute,
Mukteswar, Nainital, India
Received February 23, 2004; accepted April 28, 2004
Published online June 30, 2004
Summary. A sandwich ELISA test using PPR speciﬁc monoclonal antibody
(clone 4G6) to an epitope of nucleocapsid protein has been developed. The test
uses polyclonal sera to capture the antigen from clinical samples (swabs and
tissues). Captured antigens from clinical samples are detected using PPR speciﬁc
monoclonal antibody. The test is speciﬁc to PPR as it failed to detect rinderpest
vaccine virus (RBOK strain). Varieties of clinical samples originating from labo-
ratory experiments (n = 231) and from ﬁeld (n = 259) were employed to test the
efﬁcacy of sandwich-ELISA test. The test compared very well with an interna-
tionally accepted commercial Immune-capture ELISA kit, which uses biotinylated
monoclonal antibody against the nucleocapsid protein. On a parallel testing using
490 clinical samples, 4G6 MAb based sandwich ELISA had an overall relative
diagnostic speciﬁcity of 92.8% and diagnostic sensitivity of 88.9% compared to
the commercial kit. The newly developed test is free from prozone phenomenon.
PPR outbreaks from various parts of India have been conﬁrmed using the test.
Findings suggested that the newly developed ELISA is suitable for PPR diagnosis
under ﬁeld conditions.
Peste des petits ruminants (PPR) is an acute febrile viral disease of small rumi-
nants characterized by necrotizing and erosive stomatitis, enteritis and pneumonia
[12, 10]. It was ﬁrst described in West Africa in 1942 . In India, the disease was
ﬁrst reported from Tamilnadu in the year 1987  and became endemic in the
country [4, 18, 27, 30]. India is a vast country with a population of 126 million
goats and 58.2 million sheep. Emergence of PPR is now considered to be one of
the main constraints to improve the production of small ruminants in this country.