Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for Toxoplasma infection in pigs

The Journal of Veterinary Medical Science

article

Hieu Duc DUONG¹  Yuji TANIGUCHI³  Yasuhiro TAKASHIMA³  Satoshi SEKIGUCHI⁴  Khin Myo AYE⁶  Parnian AHMADI¹  Linh Khanh BUI²  Takao IRIE¹  Eiji NAGAYASU⁶  Ayako YOSHIDA¹ 

[1] Laboratory of Veterinary Parasitic Diseases, Department of Veterinary Sciences, Faculty of Agriculture, University of Miyazaki;Department of Veterinary Parasitology, Faculty of Veterinary Medicine, Vietnam National University of Agriculture;Department of Veterinary Parasitology, Faculty of Applied Biological Sciences, Gifu University;Laboratory of Animal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of Miyazaki;Centre for Animal Diseases Control ,(CADIC), University of Miyazaki;Division of Parasitology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki;Parasitology Research Division, Department of Medical Research

关键词: Luciferase-linked Antibody Capture Assay (LACA);    nanoluciferase;    pig;    serodiagnosis;    Toxoplasma gondii;   

DOI  :  10.1292/jvms.22-0099

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

Toxoplasmosis is a widespread protozoan zoonosis. Since ingesting undercooked meat harboring Toxoplasma gondii cyst is considered one of the major transmission routes to humans, the screening of T. gondii in meat-producing animals can reduce the risk of food-borne toxoplasmosis in humans. Among serological diagnostic methods, Luciferase-linked Antibody Capture Assay (LACA) has been found to be a promising platform with high sensitivity and specificity. In this study, we aimed to evaluate recombinant nanoluciferase fused-T. gondii antigens (rNluc-GRA6, rNluc-GRA7, rNluc-GRA8 and rNluc-BAG1) for their potential use in LACA for pigs. As a result, the sensitivity of GRA6-, GRA7-, GRA8- and BAG1-LACA were 70.0%, 80.0%, 80.0% and 30.0% with specificity 87.0%, 81.5%, 74.1% and 50.0%, respectively. The cocktail LACA using a mixture of rNluc-GRA6, rNluc-GRA7 and rNluc-GRA8 indicated higher sensitivity (90.0%) and a similar specificity (96.3%) in comparison with the commercial ELISA kit. Compared to the Dye-Test as a reference test, cocktail LACA showed strong agreement (kappa value=0.811) when we assessed pig sera collected at the slaughterhouse. In addition, we also successfully established the rapid LACA format for the detection of Toxoplasma infection in pigs (called Rapid-LACA) in which the test could be performed within 30 min. In Rapid-LACA, the protein A pre-coated/blocked plates could be preserved at −30°C, 4°C or room temperature conditions for at least two months without compromising on the quality of assay.

【 授权许可】

Unknown