Recombinant cystatin-like protein-based competition ELISA for Trichinella spiralis antibody test in multihost sera

PLoS Negl Trop Dis. 2021 Aug 25;15(8):e0009723. doi: 10.1371/journal.pntd.0009723. eCollection 2021 Aug.

Abstract

Objectives: Trichinella spiralis is a zoonotic parasite with a complex parasitic life cycle and exposed to animals or humans by infectious meat. To control transmissions of T. spiralis through the food chain to humans, sensitive and selective multihost sera-diagnosis is urgent needed for monitoring T. spiralis exposure.

Methods: A competition enzyme-linked immunosorbent assay (cELISA) for T. spiralis infection diagnosis in multihost sera was developed based on recombinant cystatin-like protein (rCLP-cELISA) as well as monoclonal antibodies. The sensitivity and accuracy of the rCLP-cELISA were quantified using swine (n = 1316), mice (n = 189) and human (n = 157) serum samples. T. spiralis-antibody targeting test ability of the rCLP-cELISA in swine (n = 22) and human (n = 36), instead of other parasites or viruses antibodies, was evaluated.

Results: The rCLP-cELISA showed high agreement with commercial ELISA kits in field swine sera assessed by Cohen's kappa value (κ = 0.7963). And it showed 100% specificity in human trichinellosis detection with sensitivity of 96.49%, no cross-reaction with other parasite or virus infections, and high positive detection rate of 87.5% in low-dose infected swine. Besides, the rCLP-cELISA exhibited potential in the detection of T. spiralis, T. nelsoni and Trichinella T8 infections.

Conclusions: The rCLP-cELISA can be used for T. spiralis-associated antibody test in multihost sera.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Helminth / blood*
  • Cystatins / genetics
  • Cystatins / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Helminth Proteins / genetics
  • Helminth Proteins / immunology
  • Humans
  • Mice
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Swine
  • Swine Diseases / blood*
  • Swine Diseases / parasitology
  • Trichinella spiralis / genetics
  • Trichinella spiralis / immunology
  • Trichinella spiralis / isolation & purification*
  • Trichinellosis / blood*
  • Trichinellosis / parasitology

Substances

  • Antibodies, Helminth
  • Cystatins
  • Helminth Proteins
  • Recombinant Proteins

Grants and funding

ML was supported by the National Key Research and Development Program of China (2018YFC1602500, 2017YFC1601200), the National Natural Science Foundation of China (NO. 31520103916), Guangdong innovation and Enterpreneurial Research Team Program (NO. 2014ZT05S123), Jilin Provincial Science and Technology Development Project (20180520042JH), and Program for JLU Science and Technology Innovative Research Team (2017TD-32), http://www.nsfc.gov. http://www.jlkjxm.com; XL prof. was supported by the National Natural Science Foundation of China (NO. 31872467), http://www.service.most.gov.cnhttp://www.nsfc.gov.; YZ prof. was supported by the National Natural Science Foundation of China (NO. 31871888), http://www.nsfc.gov. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.