Development of EMA-2 recombinant antigen based enzyme-linked immunosorbent assay for seroprevalence studies of Theileria equi infection in Indian equine population.
Equine piroplasmosis is a tick-transmitted protozoan disease caused by Theileria equi and/or Babesia caballi. In the present study, we expressed a 53kDa protein from the truncated EMA-2 gene of T. equi (Indian strain) and developed EMA-2ELISA using this expressed protein. This ELISA is able to detect T. equi-specific antibodies in experimentally infected animals as early as 9 days post-infection. The assay developed was validated with the OIE recommended competitive ELISA (cELISA) on 120 serum samples and significant agreement (kappa=0.93) was observed between results of both the ELISAs which indicates suitability of EMA-2ELISA for use in sero-diagnosis. Diagnostic sensitivity and specificity of EMA-2ELISA - as compared with cELISA - were 0.97 and 0.96, respectively. Analysis of 5651 equine serum samples - collected during 2007-2012 from 12 states of India representing eight agro-climatic zones - by EMA-2ELISA revealed 32.65% seroprevalence of T. equi in India. In conclusion, the EMA-2ELISA developed using the T. equi EMA-2 recombinant protein as antigen for detecting T. equi-specific antibodies has good diagnostic potential for sero-epidemiological surveys.
National Research Centre on Equines, Sirsa Road, Hisar 125001, Haryana, India. Electronic address: kumarsanjay66@yahoo.com.
Sanjay
S
Kumar
Rajender
R
Kumar
Ashok K
AK
Gupta
Suresh C
SC
Yadav
Sachin K
SK
Goyal
Sandip K
SK
Khurana
Raj K
RK
Singh
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D001403:Babesia / Q000145:classification / Q000235:genetics; D004247:DNA / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D018397:Equidae; D006734:Horse Diseases / Q000453:epidemiology / Q000469:parasitology*; D006736:Horses; D007194:India / Q000453:epidemiology; D008246:Lysophospholipids; D015800:Protozoan Proteins / Q000235:genetics*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies; D016794:Theileria / Q000302:isolation & purification*; D013801:Theileriasis / Q000175:diagnosis / Q000453:epidemiology / Q000469:parasitology*
C099529:1-O-(9',10'-methanohexadecanoyl)-sn-glycero-2,3-cyclic phosphate; D000953:Antigens, Protozoan; C111468:EMA-2 protein, Babesia equi; D008246:Lysophospholipids; D015800:Protozoan Proteins; D012337:RNA, Ribosomal, 18S; D011994:Recombinant Proteins; D004247:DNA
Babesia equi; ELISA; EMA-2; Equine piroplasmosis; Seroprevalence; Theileria equi