Molecular cloning and characterization of a major egg antigen in Paragonimus westermani and its use in ELISA for the immunodiagnosis of paragonimiasis.
A recombinant protein of a Paragonimus westermani egg antigen was produced and tested as an antigen for the serologic diagnosis of P. westermani infection. The P. westermani egg antigen gene contains a single open reading frame of 966 base pairs encoding 322 amino acids from 5' methionine to the 3' stop codon. The predicted amino acid sequence of this egg antigen was 40, 38, and was 35% identical to heat shock proteins from Schistosoma japonicum, Schistosoma mansoni, and Taenia saginata. The distribution this antigen was investigated in adult worms by indirect immunofluorescence assay, and found to be distributed in eggs and uteri. The specificity and sensitivity of the recombinant antigen were assessed by enzyme-linked immunosorbent assay (ELISA) using sera from patients infected with different parasites, which included 41 patients with paragonimiasis, and negative controls. The diagnostic positive and negative predictive absorbance value was 0.24 and the sensitivity of ELISA using the recombinant antigen was 90.2%, and its specificity 100%. Our results suggest that the developed recombinant major egg antigen-based ELISA offers a highly sensitive and specific assay for the diagnosis of paragonimiasis.
Lee
Jae Sik
JS
Department of Parasitology and Institute of Tropical Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, 120-752, South Korea.
Lee
Jongweon
J
Kim
Sun Hyun
SH
Yong
Tai-Soon
TS
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015723:Gene Library; D008969:Molecular Sequence Data; D010063:Ovum / Q000276:immunology*; D010237:Paragonimiasis / Q000175:diagnosis* / Q000276:immunology; D048492:Paragonimus westermani / Q000276:immunology* / Q000302:isolation & purification*
D000947:Antigens, Helminth; D016054:DNA, Protozoan