Molecular cloing and bioinformatics analysis of lactate dehydrogenase from Taenia multiceps

Parasitol Res. 2017 Oct;116(10):2845-2852. doi: 10.1007/s00436-017-5568-x. Epub 2017 Aug 1.

Abstract

Coenurus cerebralis, the larval stage (metacestode or coenurus) of Taenia multiceps, parasitizes sheep, goats, and other ruminants and causes coenurosis. In this study, we isolated and characterized complementary DNAs that encode lactate dehydrogenase A (Tm-LDHA) and B (Tm-LDHB) from the transcriptome of T. multiceps and expressed recombinant Tm-LDHB (rTm-LDHB) in Escherichia coli. Bioinformatic analysis showed that both Tm-LDH genes (LDHA and LDHB) contain a 996-bp open reading frame and encode a protein of 331 amino acids. After determination of the immunogenicity of the recombinant Tm-LDHB, an indirect enzyme-linked immunosorbent assay (ELISA) was developed for preliminary evaluation of the serodiagnostic potential of rTm-LDHB in goats. However, the rTm-LDHB-based indirect ELISA developed here exhibited specificity of only 71.42% (10/14) and sensitivity of 1:3200 in detection of goats infected with T. multiceps in the field. This study is the first to describe LDHA and LDHB of T. multiceps; meanwhile, our results indicate that rTm-LDHB is not a specific antigen candidate for immunodiagnosis of T. multiceps infection in goats.

Keywords: Bioinformatic analysis; Indirect ELISA; Lactate dehydrogenase; Taenia multiceps.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cestode Infections / diagnosis
  • Cestode Infections / parasitology
  • Cestode Infections / veterinary*
  • Cloning, Molecular
  • Computational Biology
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Goat Diseases / diagnosis
  • Goat Diseases / parasitology*
  • Goats
  • L-Lactate Dehydrogenase / chemistry
  • L-Lactate Dehydrogenase / genetics*
  • L-Lactate Dehydrogenase / isolation & purification
  • L-Lactate Dehydrogenase / metabolism
  • Larva
  • Models, Molecular
  • Molecular Conformation
  • Phylogeny
  • Sensitivity and Specificity
  • Sequence Alignment / veterinary
  • Serologic Tests / veterinary
  • Taenia / enzymology*
  • Taenia / genetics
  • Taenia / immunology
  • Taenia / isolation & purification

Substances

  • L-Lactate Dehydrogenase