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Cloning and expression of DiT33 from Dirofilaria immitis: a specific and early marker of heartworm infection

Published online by Cambridge University Press:  06 April 2009

X. Q. Hong ,
J. Santiago Mejia ,
S. Kumar ,
F. B. Perler  and
C. K. S. Carlow
X. Q. Hong
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
J. Santiago Mejia
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
S. Kumar
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
F. B. Perler
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
C. K. S. Carlow
Affiliation:
New England Biolabs, Beverly, MA 01915, USA
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Summary

Dirofilaria immitis is an important filarial parasite of dogs and cats, and a useful model for human filariasis. Current diagnostic tests for heartworm infection in animals rely on the presence of fecund female worms (usually found 6·5 months post-infection or later) and therefore fail to detect pre-patent infections. Putative pepsin inhibitors from 2 filarial parasites of humans namely Onchocerca volvulus (Ov33, Oc3.6, OvDSB) and Brugia malayi (Bm33), have been shown to be useful in diagnosis of onchocerciasis and lymphatic filariasis, respectively. Previous studies have suggested that a homologue exists in D. immitis (DiT33), which may have potential in diagnosis of heartworm infection. In this study, the isolation and characterization of a cDNA clone encoding DiT33 is described.‡ This cDNA contains 12 bases of the nematode-specific 22 nucleotide spliced leader sequence and encodes a 26·4 kDa-protein with a high level of similarity (87–89%) to other filarial members of the family. DJT33 was over-expressed in E. coli as a fusion with the maltose-binding protein and serological analysis was performed using a panel of clinically defined dog sera. The findings of this study indicate that DiT33 is a promising antigen for the early detection of D. immitis and may be a valuable tool in the control and management of heartworm infection.

Keywords

Dirofilaria immitisDiT33Ov33pepsin inhibitordiagnosisheartworm.
Type
Research Article
Information
Parasitology , Volume 112 , Issue 3 , March 1996 , pp. 331 - 338
Copyright
Copyright © Cambridge University Press 1996

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