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CLONING AND CHARACTERIZATION OF A NEW CYSTEINE PROTEINASE SECRETED BY PARAGONIMUS WESTERMANI ADULT WORMS
- Source :
- The American Journal of Tropical Medicine and Hygiene. 71:87-92
- Publication Year :
- 2004
- Publisher :
- American Society of Tropical Medicine and Hygiene, 2004.
-
Abstract
- The cysteine proteinases of Paragonimus westermani are known to play important roles in invasion and pathogenesis to hosts and in immune modulation and nutrient uptake. In this study, we have cloned a new cysteine proteinase of P. westermani, PwCP2, from adult worms and tested its diagnostic usefulness. The PwCP2 gene had an open reading frame of 816 base pairs and a conserved catalytic triad of cysteine, histidine, and asparagine residues. The mature form of recombinant PwCP2 (rPwCP2) lacking a proregion was overexpressed in Escherichia coli and used to produce antiserum. Western blot and immunohistochemical analyses using this antiserum showed that PwCP2 was expressed as a mature form, 24-kD product in a crude extract and in the excretory-secretory product of P. westermani, and was localized mainly in the intestinal epithelium of the adult worm. Western blot analysis using the rPwCP2 showed not only high sensitivity (90%) and specificity (100%) to sera from patients with paragonimiasis westermani, but also no cross-reactivity with sera from patients with clonorchiasis, sparganosis, or cysticercosis. Furthermore, an enzyme-linked immunosorbent assay using rPwCP2 exhibited a sensitivity of 93% and a specificity of 93% with sera of rats infected with P. westermani metacercariae. These results suggest that the excretory-secretory PwCP2 can be used for the diagnosis of paragonimiasis.
- Subjects :
- Paragonimus westermani
Paragonimiasis
Molecular Sequence Data
Paragonimus
Enzyme-Linked Immunosorbent Assay
Aspergillopepsin II
Sensitivity and Specificity
Substrate Specificity
Microbiology
law.invention
Western blot
law
Virology
parasitic diseases
medicine
Animals
Amino Acid Sequence
Cloning, Molecular
Antiserum
biology
medicine.diagnostic_test
biology.organism_classification
medicine.disease
Recombinant Proteins
Rats
Cysteine Endopeptidases
Infectious Diseases
Clonorchiasis
Recombinant DNA
Parasitology
Cysteine
Subjects
Details
- ISSN :
- 14761645 and 00029637
- Volume :
- 71
- Database :
- OpenAIRE
- Journal :
- The American Journal of Tropical Medicine and Hygiene
- Accession number :
- edsair.doi.dedup.....c70ce46f11b92a0cb38d4108ef4d0687