Development and Evaluation of an Enzyme-Linked Immunosorbent Assay with Recombinant SAG2 for Diagnosis of Toxoplasma gondii Infection in Cats

Huang, Xiaohong; Xu, Xuan; Kimbita, Elikira; Battur, Banzragch; Miyazawa, Takayuki; Fukumoto, Shiya; Mishima, Masayuki; Makala, Levi; Suzuki, Hiroshi; Sugimoto, Chihiro; Nagasawa, Hideyuki; Fujisaki, Kozo; Mikami, Takeshi; Igarashi, Ikuo

doi:10.1645/0022-3395(2002)088[0804:daeoae]2.0.co;2

Cited by 31 publications

(26 citation statements)

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“…The latex agglutination test (LAT) is a commercially available test for the diagnosis of feline toxoplasmosis. In previous studies, the surface antigen 2 (SAG2) of T. gondii expressed either in Escherichia coli or in insect cells was validated as a useful antigen which promised a highly sensitive and specific enzyme-linked immunosorbent assay (ELISA) (6,7). However, the performance of ELISA, which is laborious and time-consuming and requires expertise and equipment, still remains to be improved.…”

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confidence: 99%

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Rapid Immunochromatographic Test Using Recombinant SAG2 for Detection of Antibodies against Toxoplasma gondii in Cats

Huang

Hirata

et al. 2004

J Clin Microbiol

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confidence: 99%

“…2. LAT and ELISA were performed as described previously (6). Relative sensitivity, specificity, and agreement with LAT and ELISA were calculated as described by Griner et al (5).…”

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confidence: 99%

Rapid Immunochromatographic Test Using Recombinant SAG2 for Detection of Antibodies against Toxoplasma gondii in Cats

Huang

Hirata

et al. 2004

J Clin Microbiol

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“…SAG1antigen has also been shown to be effective in the sero-diagnosis in cats (Huang et al 2002;Kimbita et al 2001). This antigen is stage specific antigen that only exists in tachyzoite stage of T. gondii (Kasper et al 1984).…”

Section: Discussionmentioning

confidence: 99%

Seroprevalence of Toxoplasma gondii infection in domestic dogs in an area from northwest of Iran: a cross-sectional study using immunodominant surface antigen 1 (SAG1)

et al. 2015

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Toxoplasma gondii is an obligate intracellular protozoan parasite that infects humans and animals. T. gondii surface antigen 1 (SAG1) is an appropriate antigen with high specificity and sensitivity for the detection of T. gondii infection in humans and animal hosts. The aim of this study was to determine the seroprevalence of T. gondii infection using SAG1 antigen (P30) in ownership dogs in Meshkin-Shahr district in the northwestern Iran. The sera samples were collected from 171 domestic dogs and tested using indirect ELISA (SAG1 antigen). The data were analyzed using SPSS software version 13. From a total of 171 dogs, 82 (48 %) of them were sero-positive. No statistical significant difference was seen between T. gondii infection and gender (P = 0.995). The highest seroprevalence of rate was observed in [5 years animals; but no statistical significant difference was seen between T. gondii infection and age (P = 0.589). Our findings indicate that Toxoplasma seropositivity rate is high in ownership dogs in northwest of Iran. This is probably due to high exposure to contaminated food, soil, or water sources with sporulated Toxoplasma oocysts.

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“…The E-rSAG2 antigen was used as an antigen in an ELISA to test the same cat serum samples in our previous study (9). Comparison of the results between the two ELISAs showed that there was a significant correlation either between the OD values ( ϭ 0.92, n ϭ 183, P Ͻ 0.001) or between the titers ( ϭ 0.71, n ϭ 53, P Ͻ 0.001).…”

Section: Diagnosis Of T Gondii Infection In Cats By Elisa With Srsagmentioning

confidence: 89%

“…The recombinant SAG2 expressed in Escherichia coli (E-rSAG2) was effective in detecting the immunoglobulin G (IgG) antibody to T. gondii in human patients with acute toxoplasmosis (21,23). In our previous study, E-rSAG2 was used as an antigen for ELISA to detect T. gondii infection in domestic cats and was shown to be a good reagent for the diagnosis of toxoplasmosis (9). Vaccination with E-rSAG2 provided partial protection against a lethal infection of T. gondii and inhibited cyst formation in the brains of infected mice (16).…”

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confidence: 99%

Characterization ofToxoplasma gondiiSAG2 Expressed in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

Huang

Suzuki

et al. 2002

Clin Vaccine Immunol

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A baculovirus carrying the SAG2 gene of Toxoplasma gondii was constructed, and recombinant SAG2 protein (S-rSAG2) was expressed in insect cells. S-rSAG2 was recognized by sera from cats and pigs infected with T. gondii. Mice immunized with S-rSAG2 produced high titers of specific immunoglobulin G2a (IgG2a) and IgG1 antibodies. In an indirect fluorescent antibody test, all mouse antisera against S-rSAG2 reacted strongly to the natural parasites, but those against rSAG2 expressed in Escherichia coli (E-rSAG2) only showed very weak reaction, although no markedly difference was found in the reaction to denatured antigen, T. gondii lysate, in Western blot analysis. The results suggest that S-rSAG2 is better than E-rSAG2 in both antigenicity and immunogenicity. Enzyme-linked immunosorbent assay (ELISA) with S-rSAG2 could differentiate clearly between sera from 30 specific-pathogen-free cats and 4 experimentally infected cats. Serum samples from domestic cats in Japan were tested by the ELISA and compared with a latex agglutination test (LAT) and ELISA with E-rSAG2. Of 187 samples, all 35 LAT-positive sera had strong reactions to S-rSAG2 and E-rSAG2. Of the 152 LAT-negative sera, 18 were positive in the ELISA with S-rSAG2, whereas only 2 were positive in the ELISA with E-rSAG2. Although there were significant correlations among the three methods, the ELISA with S-rSAG2 was more sensitive than the others, which could be attributed to the fact that S-rSAG2 shares some common conformational structure with the native antigen. The results suggest that S-rSAG2 would be a useful reagent for the detection of T. gondii infection in cats.Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii. As many as one-third of the world's population shows serological evidence of infection (10). Although it is generally asymptomatic in healthy adults, it may cause abortion, neonatal death, severe sequelae in neonates, and lifethreatening lesions in AIDS patients. Furthermore, it may complicate immunosuppressive therapy. Of all the encephalitis occurring in human immunodeficiency virus-positive patients, ca. 50% is due to T. gondii infection (4). In addition to being as major source of infection for humans, it is also of considerable importance in domestic animals and is responsible for abortion in sheep and swine (25). Therefore, there is an urgent need to develop an effective diagnostic kit and vaccine.Surface antigen 2 (SAG2 and P22) of T. gondii is a major surface protein known as an attachment ligand (8) that also has good antigenicity and immunogenicity (1, 14, 21, 23). The recombinant SAG2 expressed in Escherichia coli (E-rSAG2) was effective in detecting the immunoglobulin G (IgG) antibody to T. gondii in human patients with acute toxoplasmosis (21, 23). In our previous study, E-rSAG2 was used as an antigen for ELISA to detect T. gondii infection in domestic cats and was shown to be a good reagent for the diagnosis of toxoplasmosis (9). Vaccination with E-rSAG2 provided partial protection against a lethal infection of T. g...

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Development and Evaluation of an Enzyme-Linked Immunosorbent Assay with Recombinant SAG2 for Diagnosis of Toxoplasma gondii Infection in Cats

Cited by 31 publications

References 11 publications

Rapid Immunochromatographic Test Using Recombinant SAG2 for Detection of Antibodies against Toxoplasma gondii in Cats

Rapid Immunochromatographic Test Using Recombinant SAG2 for Detection of Antibodies against Toxoplasma gondii in Cats

Seroprevalence of Toxoplasma gondii infection in domestic dogs in an area from northwest of Iran: a cross-sectional study using immunodominant surface antigen 1 (SAG1)

Characterization ofToxoplasma gondiiSAG2 Expressed in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

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