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. 2014 Feb;52(1):41-6.
doi: 10.3347/kjp.2014.52.1.41. Epub 2014 Feb 19.

Diagnostic efficacy of a recombinant cysteine protease of Spirometra erinacei larvae for serodiagnosis of sparganosis

S M Mazidur Rahman  1 , Jae-Hwan Kim  1 , Sung-Tae Hong  1 , Min-Ho Choi  1
Affiliations

Diagnostic efficacy of a recombinant cysteine protease of Spirometra erinacei larvae for serodiagnosis of sparganosis

S M Mazidur Rahman et al. Korean J Parasitol. 2014 Feb.

Abstract

The mature domain of a cysteine protease of Spirometra erinacei plerocercoid larva (i.e., sparganum) was expressed in Escherichia coli, and its value as an antigen for the serodiagnosis of sparganosis was investigated. The recombinant protein (rSepCp-1) has the molecular weight of 23.4 kDa, and strongly reacted with the sparganum positive human or mice sera but not with negative sera by immunoblotting. ELISA with rSepCp-1 protein or sparganum crude antigen (SeC) was evaluated for the serodiagnosis of sparganosis using patient's sera. The sensitivity and specificity of ELISA using rSepCp-1 protein were 95.0% (19/20) and 99.1% (111/112), respectively. In contrast, the sensitivity and specificity of ELISA with SeC were 100% (20/20) and 96.4% (108/112), respectively. Moreover, in experimentally infected mice, the sensitivity and specificity of both ELISA assays were 100% for the detection of anti-sparganum IgG. It is suggested that the rSepCp-1 protein-based ELISA could provide a highly sensitive and specific assay for the diagnosis of sparganosis.

Keywords: ELISA; Spirometra erinacei; cysteine protease; serodiagnosis; sparganum.

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Conflict of interest statement

We have no conflict of interest related with this work.

Figures

Fig. 1
Fig. 1
SDS-PAGE and immunoblot analysis of recombinant Spirometra erinacei plerocercoid cysteine protease (rSepCp-1). (A) Histidine-tagged rSepCp-1 protein visualized by SDS-PAGE and Coomassie blue staining. Lane M, molecular weight marker; lane 1, E. coli lysate without isopropyl-β-D-thiogalactopyranoside (IPTG) induction; lane 2, E. coli lysate with IPTG induction; lane 3, rSepCp-1 protein purified by Ni-NTA affinity chromatography. (B) Immunoblot analysis of the same protein samples as in 'A' using a mouse anti-histidine monoclonal antibody. (C) Immunoblotting of the rSepCp-1 protein with sparganum-negative (lane 1) and sparganum-positive (lane 2) human serum samples. (D) Immunoblotting of the rSepCp-1 protein with sparganum-negative (lane 1) and sparganum-positive (lane 2) mouse serum samples.
Fig. 2
Fig. 2
Comparison of the absorbances of the ELISA using the crude antigen (A) and the ELISA using the recombinant Spirometra erinacei plerocercoid cysteine protease (rSepCp-1) (B) for the detection of anti-sparganum IgG in human serum samples. The cut-off values are indicated by the dashed horizontal lines. Sp, sparganosis; H, healthy control; Toxo, toxocariasis; Cs, clonorchiasis; Cys, cysticercosis; PW, paragonimiasis; An, anisakiasis.
Fig. 3
Fig. 3
Comparison of absorbances by ELISA using the crude antigen (A) and ELISA using the recombinant Spirometra erinacei plerocercoid cysteine protease (rSepCp-1) (B) for the detection of anti-sparganum IgG in serum samples of infected and uninfected mice. The cut-off values are indicated by the dashed horizontal lines.
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