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Expression of biologically active recombinant gal/galnac specific lectin of entamoeba histolytica and diagnostic test for e. histolytica infection in preserved fecal samples
Expression of biologically active recombinant gal/galnac specific lectin of entamoeba histolytica and diagnostic test for e. histolytica infection in preserved fecal samples
Invasive amoebiasis is caused by the pathogenic protozoa iEntamoeba histolytica/i. iE. histolytica/i is genetically distinct but morphologically identical to a nonpathogenic species iEntamoeba dispar/i. Current methods for distinguishing between these species generally require the use of fresh, unpreserved stool specimens. This requirement limits their use in developed countries since the great majority of specimens are submitted preserved in stool fixative. This invention provides a method for generating monoclonal antibodies against the recombinant 170 kDa subunit of the Gal/GalNAc specific lectin of iE. histolytica/i, that specifically recognize iE. histolytica/i, but not iE. dispar/i, in preserved stool samples. These antibodies do not cross-react with other bowel protozoa including iEntamoeba coli, Giardia lamblia/i, and iDientamoeba fragilis/i. This invention also provides a method of expressing a functional recombinant 170 kDa subunit of the Gal/GalNAc specific lectin of iE. histolyca/i and its cysteine-rich fragment in iSpodoptera frugiperda/i cells infected by recombinant baculoviruses: The recombinant protein is used for developing improved vaccines and diagnostic tests.
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