An ELISA using recombinant TmHSP70 for the diagnosis ofTaenia multiceps
infections in goats
Yu Wang,Huaming Nie,Xiaobin Gu, Tao Wang, XingHuang, Lin Chen, Weimin Lai, Xuerong Peng, Guangyou Yang
Abstract
Infections with the tapewormTaenia multicepsare problematic for ruminant farming worldwide. Here we develop a novel and rapid method for serodiagnosis ofT. multicepsinfections via an indirect ELISA (iELISA) that uses a heat shock protein, namely, TmHSP70. We extracted the total RNA ofT. multicepsfrom the protoscoleces of cysts dissected from the brains of infected goats. Subsequently, we successfully amplified, cloned and expressed the TmHSP70 gene inEscherichia coliBL21 (DE3). Western blot analysis showed that the recombinant protein (∼34 kDa molecular weight) was recognized by the coenurosis positive serum. Given these initial, robust immunogenic properties for recombinant TmHSP protein, we assessed the ELISA-based serodiagnostic potential of this gene. The indirect ELISA was then optimized to 2.70μg/well dilution for antigen and 1:80 dilution for serum,while the cut-off value is 0.446. We report that our novel TmHSP ELISA detectedT. multicepssera with a sensitivity of 1:10240 and a specificity of 83.3% (5/6). In a preliminary application, this assay correctly confirmedT. multicepsinfection in 30 infected goats, consistent with the clinical examination. This study has revealed that our novel iELISA, which uses the rTmHSP protein, provides a rapid test for diagnosing coenurosis.
Key words
Tapeworm;Taenia multiceps; TmHSP 70; Clone; Expression; Immunodiagnosis; Indirect ELISA
Veterinary Parasitology, 2015,212:469-472. doi:10.1016/j.vetpar.2015.06.020
Read full text: http://www.sciencedirect.com/science/article/pii/S0304401715003015
