Evaluation of a Recombinant Antigen Enzyme-Linked Immunosorbent Assay (ELISA) in the Diagnostics of Antinuclear Antibodies (ANA) in Children with Rheumatic Disorders

ABSTRACT We produced a recombinant cysteine proteinase of Clonorchis sinensis and tested its value as an antigen for serologic diagnosis of C. sinensis infections. The predicted amino acid sequence of the cysteine proteinase of C. sinensis was 58, 48, and 40% identical to those of cathepsin L cysteine proteinases from Paragonimus westermani, Schistosoma japonicum, and Fasciola hepatica, respectively. Western blotting analysis showed that sera from patients infected with C. sinensis strongly reacted with the recombinant protein and that sera from patients infected with S. japonicum weakly reacted with the recombinant protein. Antibody against the recombinant protein stained proteins migrating at about 37 and 28 kDa in C. sinensis adult worm crude extracts. Immunostaining revealed that the cysteine proteinase of C. sinensis was located in the intestinal epithelial cells of the adult parasite and in intrauterine eggs. The specificity and sensitivity of the recombinant antigen or C. sinensis adult worm crude extracts were assessed by an enzyme-linked immunosorbent assay (ELISA) using serum samples from humans infected with different parasites, including 50 patients with clonorchiasis, and negative controls. The sensitivities of the ELISA with the recombinant antigen and C. sinensis adult worm crude extracts were 96 and 88%, respectively. The specificities of the ELISA with the recombinant antigen and C. sinensis adult worm crude extracts were 96.2 and 100%, respectively. The results suggested that the recombinant cysteine proteinase-based ELISA could provide a highly sensitive and specific assay for diagnosis of clonorchiasis.

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Evaluation of Babesia bigemina 200 kDa recombinant antigen in enzyme-linked immunosorbent assay

Parasitology Research ◽

10.1007/s00436-009-1392-2 ◽

2009 ◽

Vol 105 (1) ◽

pp. 249-254 ◽

Cited By ~ 4

Author(s):

Khukhuu Altangerel ◽

Andy Alhassan ◽

Hiroshi Iseki ◽

Thillaiampalam Sivakumar ◽

Damdinsuren Boldbaatar ◽

...

Keyword(s):

Immunosorbent Assay ◽

Recombinant Antigen ◽

Enzyme Linked Immunosorbent Assay ◽

Babesia Bigemina

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Recombinant antigen-based avidin-biotin microtiter enzyme-linked immunosorbent assay for serodiagnosis of invasive amebiasis.

Journal of Clinical Microbiology ◽

10.1128/jcm.34.4.828-833.1996 ◽

1996 ◽

Vol 34 (4) ◽

pp. 828-833 ◽

Cited By ~ 15

Author(s):

B R Shenai ◽

B L Komalam ◽

A S Arvind ◽

P R Krishnaswamy ◽

P V Rao

Keyword(s):

Immunosorbent Assay ◽

Recombinant Antigen ◽

Enzyme Linked Immunosorbent Assay

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Evaluation of Enzyme-Linked Immunosorbent Assay for Diagnosis of Post-Kala-Azar Dermal Leishmaniasis with Crude or Recombinant k39 Antigen

Clinical and Vaccine Immunology ◽

10.1128/cdli.9.2.370-373.2002 ◽

2002 ◽

Vol 9 (2) ◽

pp. 370-373 ◽

Cited By ~ 1

Author(s):

P. Salotra ◽

G. Sreenivas ◽

A. A. Nasim ◽

B. V. Subba Raju ◽

V. Ramesh

Keyword(s):

Immunosorbent Assay ◽

Leishmania Donovani ◽

Skin Lesions ◽

Elisa Test ◽

Recombinant Antigen ◽

Enzyme Linked Immunosorbent Assay ◽

Invasive Test ◽

Kala Azar ◽

Axenic Amastigotes ◽

Higher Sensitivity

ABSTRACT The diagnosis of post-kala-azar dermal leishmaniasis (PKDL), a dermatosis that provides the only known reservoir for the parasite Leishmania donovani in India, remains a problem. Timely recognition and treatment of PKDL would contribute significantly to the control of kala-azar. We evaluated here the potential of the enzyme-linked immunosorbent assay (ELISA) as a diagnostic tool for PKDL. Antigen prepared from promastigotes and axenic amastigotes with parasite isolates that were derived from skin lesions of a PKDL patient gave sensitivities of 86.36 and 92%, respectively, in the 88 PKDL cases examined. The specificity of the ELISA test was examined by testing groups of patients with other skin disorders (leprosy and vitiligo) or coendemic infections (malaria and tuberculosis), as well as healthy controls from areas where this disease is endemic or is not endemic. A false-positive reaction was obtained in 14 of 144 (9.8%) of the controls with the promastigote antigen and in 14 of 145 (9.7%) of the controls with the amastigote antigen. Evaluation of the serodiagnostic potential of recombinant k39 by ELISA revealed a higher sensitivity (94.5%) and specificity (93.7%) compared to the other two antigens used. The data demonstrate that ELISA with crude or recombinant antigen k39 provides a relatively simple and less-invasive test for the reliable diagnosis of PKDL.

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