rTES-30USM: cloning via assembly PCR, expression, and evaluation of usefulness in the detection of toxocariasis
| Autor: | M. Suharni, Josef S. B. Tuda, N. Rahmah, A. Norhaida, A. T. Liza Sharmini |
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| Rok vydání: | 2008 |
| Předmět: |
Male
Molecular Sequence Data Helminthiasis Enzyme-Linked Immunosorbent Assay Helminth genetics Polymerase Chain Reaction Sensitivity and Specificity law.invention Serology Affinity chromatography Antigen law medicine Animals Humans Serologic Tests Polymerase chain reaction Toxocariasis biology Toxocara canis Helminth Proteins biology.organism_classification medicine.disease Virology Treatment Outcome Infectious Diseases Antigens Helminth Immunology Larva Migrans Visceral Female Parasitology Targeted Gene Repair |
| Zdroj: | Annals of Tropical Medicine & Parasitology. 102:151-160 |
| ISSN: | 1364-8594 0003-4983 |
| DOI: | 10.1179/136485908x252250 |
| Popis: | Currently, the laboratory diagnosis of toxocariasis, caused by Toxocara canis or T. cati, mainly relies on serological tests. Unfortunately, however, the specificities of most of the commercial tests that are available for the serodiagnosis of this disease are not very high and this may cause problems, especially in tropical countries where co-infections with other helminths are common. In an effort to develop a serological assay with improved specificity for the detection of Toxocara infection, an IgG(4)-ELISA based on a recombinant version (rTES-30USM) of the 30-kDa Toxocara excretory-secretory antigen (TES-30) has recently been developed. To produce the antigen, the TES-30 gene was cloned via assembly PCR, subcloned into a His-tagged prokaryotic expression vector, and purified by affinity chromatography using Ni(2+)-nitrilotriacetic-acid (Ni-NTA) resin. The performance of the ELISA based on the recombinant antigen was then compared with that of commercial kit, based on an IgG-ELISA, for the serodiagnosis of toxocariasis (Toxocara IgG-ELISA; Cypress Diagnostics, Langdorp, Belgium). Both assays were used to test 338 serum samples, including 26 samples from probable cases of toxocariasis. Assuming that all the probable cases were true cases, the assay based on rTES-30USM demonstrated a sensitivity of 92.3% (24/26) and a specificity of 89.6% (103/115) whereas the commercial kit exhibited a sensitivity of 100% (26/26) but a specificity of only 55.7% (64/115). The high sensitivity and specificity exhibited by the new IgG(4)-ELISA should make the assay a good choice for use in tropical countries and any other area where potentially cross-reactive helminthic infections are common. |
| Databáze: | OpenAIRE |
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