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Studies on the development of a DIVA assay for Haemorrhagic Septicaemia

KrishiKosh

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Title Studies on the development of a DIVA assay for Haemorrhagic Septicaemia
 
Creator Qureshi, Sabia
 
Contributor Saxena, H. M.
 
Subject P. multocida
Phage
IROMPs
DIVA
marker vaccine
alum precipitated vaccine
MAT
IHA
Haemorrahagic septicaemia
ELISA
 
Description The present study was planned to explore the possibility of development of a marker vaccine against H&S and a DIVA assay to differentiate between vaccinated and infected animals under field conditions. A novel phage lysate marker vaccine prepared by growing P. multocida (B:2) under iron restricted conditions was given to rabbits, mice and cattle. Humoral immune response to the marker and conventional alum precipitated HS vaccines were monitored using MAT, IHA and ELISA. Consistently higher antibody titres in marker vaccine groups was observed at all the stages of immunization. The antibody levels in the marker vaccine group at 240 and 210 DPI, respectively were significantly higher than those of conventional vaccine group in mice and cattle. The maximum MAT log10 titre (1.95) in case of marker vaccinated rabbits was achieved at 30DPI while the corresponding titre of 1.50 in conventional vaccinated rabbits was achieved at 90DPI. Mean log10 titres by ELISA in marker vaccinated and conventional vaccinated rabbits were 2.43 and 2.35 respectively at 30DPI whereas at 120DPI the titres in marker and conventional vaccinated rabbits were 3.29 and 2.75 for marker and conventional vaccinated rabbits, respectively. The mean antibody titre in case of marker vaccinated mice ranged from 2.59-1.73 during the period between 90-240DPI whereas the corresponding titres in conventional vaccinated mice were between 1.39-0.42 respectively, by MAT. The peak log 10 values (3.46) in case of marker vaccinated mice estimated by ELISA were attained at 90DPI whereas in conventional vaccinated mice the peak values at 90DPI were 2.82. In case of marker vaccinated cattle antibody titres (log10) peaked to 2.204 and 2.279 by MAT and IHA, respectively at 60 DPI whereas in conventional vaccinated cattle titres of 2.13 and 2.05 were observed at 60DPI by MAT and IHA respectively. Mean log10 titres by ELISA in marker vaccinated and conventional vaccinated cattle groups were 2.33 and 1.70 respectively at 90DPI whereas at 120DPI the titres in marker and conventional vaccinated cattle were 2.12 and 1.57 respectively.The immunoblot analysis of marker vaccinated animals revealed the presence of 137KDa IROMP which was absent in case of blots probed with sera of conventional HSAV vaccinated or infected animals thus validating the DIVA hypothesis. The DIVA ELISA based on antibody to 137KDa IROMP antigen could identify the marker vaccinated animals having the antibody which was absent in the conventional vaccinated and infected animals.
 
Date 2016-08-16T15:16:00Z
2016-08-16T15:16:00Z
2014-06-26
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/72636
 
Language en
 
Format application/pdf