The invention provides a neosporosis rELISA
antibody assay kit. The neosporosis
recombinant enzyme-linked immunosorbent
assay (rELISA)
antibody assay kit is characterized in that designing a primer set according to a neospora caninum NcSRS2
gene of the
GenBank, introducing the primer set to an
enzyme digestion site, removing a
signal peptide zone of an NcSRS2
protein, extracting
a DNA from Xinjiang cow positive serum, carrying out
polymerase chain reaction (PCR) amplification of the
DNA to obtain a tNcSRS2 genetic fragment having length of 975bp,
cloning the tNcSRS2 genetic fragment into a pMD-18-T carrier, carrying out
enzyme digestion and sequencing, further directionally connecting the desired fragment obtained by the previous step to a pGEX-4T-2 expression carrier, transferring the pGEX-4T-2 expression carrier with the desired fragment into
escherichia coli, carrying out inducible expression and purification, wherein expressed
fusion protein molecular weight is about 62kDa, and carrying out ELISA
antigen coating by the expressed
fusion protein to obtain the neosporosis rELISA
antibody assay kit. A use method of the neosporosis rELISA antibody assay kit comprises the following steps that a coated
enzyme label reaction plate is taken out, is subjected to first washing, is enclosed, is subjected to second washing, is added with a sample needing to be detected, is subjected to third washing, is added with a secondary antibody, is subjected to fourth washing, and undergoes a
color reaction; and when the
color reaction is finished, an OD450 / 630 value of the coated enzyme
label reaction plate is read and a result is determined. Compared with a commercial
ELISA assay kit, the neosporosis rELISA antibody assay kit has a higher
coincidence rate above 95% The neosporosis rELISA antibody assay kit also is suitable for the ELISA of a neospora caninum antibody in
ruminant serum and
blood plasma.