ELISA based on a recombinant Paragonimus heterotremus protein for serodiagnosis of human paragonimiasis in Thailand

Parasites & Vectors
Kanokkarn PothongParon Dekumyoy

Abstract

Paragonimus heterotremus is the main causative agent of paragonimiasis in Thailand. In Western blot diagnostic assays for paragonimiasis, the 35 kDa band present in crude P. heterotremus somatic extracts represents one of the known diagnostic bands. This study aimed to use a P. heterotremus cDNA library to create a recombinant version of this antigen for use in immunodiagnosis of paragonimiasis. To accomplish this aim a cDNA expression library was constructed from adult worm mRNA and immuno-screened using antibodies from mice that had been immunized with the 35 kDa antigen. Screening resulted in the identification of an immunoreactive protein encoded by clone CE3, which contained an inserted sequence composed of 1292 base pairs. This clone was selected for use in the construction of a recombinant P. heterotremus protein because of its similarity to proactivator polypeptide. For recombinant protein expression, the CE3 gene sequence was inserted into the plasmid vector pRset and the resulting product had the expected molecular weight of 35 kDa. An IgG-ELISA based on the CE3 recombinant protein was evaluated by using sera from healthy individuals, from patients with paragonimiasis and other parasitic infections. This ELISA was per...Continue Reading

References

Jul 1, 1991·International Journal for Parasitology·I IndrawatiY Ruangkunaporn
Jul 17, 1998·Clinical and Diagnostic Laboratory Immunology·Y KongS Y Cho
Aug 26, 1998·The American Journal of Tropical Medicine and Hygiene·T Ikeda
Dec 26, 2007·Indian Journal of Ophthalmology·Rajul ParikhRavi Thomas
Jul 22, 2008·Pediatric Neurology·Zhi ChenHua Feng

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Methods Mentioned

BETA
PCR
ELISAs
ELISA

Software Mentioned

SPSS
BLASX
MedCal
Blastx
DNAMan

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