Development and Clinical Evaluation of a Rapid Serodiagnostic Test for Toxoplasmosis of Cats Using Recombinant SAG1 Antigen

Chong, Chom-Kyu; Jeong, Wooseog; Kim, Hee Young; An, Dong-Jun; Jeoung, Hye-Young; Ryu, Jeong-Eun; Ko, Ara; Kim, Yong Joo; Hong, Sung-Jong; Yang, Zhaoshou; Nam, Ho-Woo

doi:10.3347/kjp.2011.49.3.207

Cited by 22 publications

(16 citation statements)

References 23 publications

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“…Four of the recombinant antigens (SAG1, SAG2, GRA3, and GRA6) expressed in E. coli have been used in an RDT to detect infection in cats. Among them, recombinant SAG1 demonstrated the highest sensitivity and specificity, i.e., 100% and 99.2%, respectively [40]. In another study, the N-terminal half of SAG1Awas linked to the intrinsic unstructured domain of GRA2 and conjugated with 40-nm colloidal gold particles [36].…”

Section: Lateral Flow Assaymentioning

confidence: 99%

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

Khan

Noordin

2019

Eur J Clin Microbiol Infect Dis

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Infection by Toxoplasma gondii is prevalent worldwide. The parasite can infect a broad spectrum of vertebrate hosts, but infection of fetuses and immunocompromised patients is of particular concern. Easy-to-perform, robust, and highly sensitive and specific methods to detect Toxoplasma infection are important for the treatment and management of patients. Rapid diagnostic methods that do not sacrifice the accuracy of the assay and give reproducible results in a short time are highly desirable. In this context, rapid diagnostic tests (RDTs), especially with point-of-care (POC) features, are promising diagnostic methods in clinical microbiology laboratories, especially in areas with minimal laboratory facilities. More advanced methods using microfluidics and sensor technology will be the future trend. In this review, we discuss serological and molecular-based rapid diagnostic tests for detecting Toxoplasma infection in humans as well as animals.

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Section: Lateral Flow Assaymentioning

confidence: 99%

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

Khan

Noordin

2019

Eur J Clin Microbiol Infect Dis

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“…Sera were tested for the presence of anti- T . gondii IgG using a commercial ELISA kit (ID Screen Toxoplasmosis Indirect ELISA, IDVET, Montpellier, France),validated for ruminants, with known sensitivity and specificity of respectively 91.3% and 98.7% [ 40 ]. The validation study of the ELISA test was previously conducted by the manufacturer (IDVET, Montpellier, France) providing values of the coefficient of variation (CV%).…”

Section: Methodsmentioning

confidence: 99%

Toxoplasma gondii Infection in Alpine Red Deer (Cervus elaphus): Its Spread and Effects on Fertility

Formenti

Trogu

Pedrotti³

et al. 2015

PLoS ONE

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In contrast to the depth of knowledge on the pathological effects of parasitism in domestic animals, the impact of the vast majority of parasites on wildlife hosts is poorly understood and, besides, information from domestics is rarely usable to disclose the parasites’ impact on free-ranging populations’ dynamics. Here we use Toxoplasmosis as a study model since, until now, the infection process and the protozoan’s effects in natural conditions has received little attention. We analysed 81 sera from red deer (Cervus elaphus) sampled in Italian Alps and through generalized linear models we evaluated (1) the epidemiological factors influencing T. gondii infection dynamics; (2) its impact on female fertility. High seroprevalence of T. gondii infection was recorded in yearling (1 year-old; prevalence = 52.4%) and adult (>2 year-old; prevalence = 51.3%) red deer, while calves (<1 year-old) did not contract the infection suggesting horizontal transmission as the main route of infection. The stable prevalence between yearlings and adults and the higher serological titres of younger individuals lead to two alternative infection processes suggesting a difference between age classes or in acquiring the infection or in responding to the pathogen. No associations between T. gondii serological titres and pregnancy status was observed indicating no direct effect on the probability of being pregnant; nevertheless a relation between females’ higher serological titres and lower foetal development emerged, suggesting potential effects of the parasite infection on deer reproduction. The results demonstrate high seroprevalence of T. gondii infection in free-ranging red deer and, furthermore, the effect on foetal development suggests the potential impact of the parasite on red deer fertility and thus on its population dynamics.

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“…In contrast, we applied a bacterially expressed biotinylated recombinant TgSAG1 [37], the major surface antigen of T. gondii tachyzoites [24], which is used widely for serodiagnosis in humans, but also in wild or livestock animals (e.g. [47][48][49][50][51][52][53][54][55]). Our recombinant TgSAG1 bio is unique in that it allows an oriented and reproducible coupling of the antigen to streptavidin-coated magnetic Luminex beads via a single C-terminal biotin [37].…”

Section: Discussionmentioning

confidence: 99%

Fluorescent bead-based serological detection of Toxoplasma gondii infection in chickens

Fabian

Hedar

Koethe

et al. 2020

Preprint

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Background: Free-ranging chickens are often infected with Toxoplasma gondii and seroconvert upon infection. This indicates environmental contamination with T. gondii. Methods: Here, we established a bead-based multiplex assay (BBMA) using the Luminex technology for the detection of T. gondii infections in chickens. Recombinant biotinylated T. gondii surface antigen 1 (TgSAG1bio) bound to streptavidin-conjugated magnetic Luminex beads served as antigen. Serum antibodies were detected by a fluorophore-coupled secondary antibody. Beads of differing color codes were conjugated with anti-chicken IgY or chicken serum albumin and served for each sample as an internal positive or negative control, respectively. The assay was validated with sera from experimentally and naturally infected chickens. The results were compared to those from reference methods, including other serological tests, PCRs and bioassay in mice.Results: In experimentally infected chickens, the vast majority (98.5%, n = 65/66) of birds tested seropositive in the BBMA. This included all chickens positive by magnetic-capture PCR (100%, n = 45/45). Most, but not all inoculated and TgSAG1bio-BBMA-positive chickens were also positive in two previously established TgSAG1-ELISAs (TgSAG1-ELISASL, n = 61/65; or TgSAG1-ELISASH, n = 60/65), or positive in an immunofluorescence assay (IFAT, n = 64/65)) and in a modified agglutination test (MAT, n = 61/65). All non-inoculated control animals (n = 28/28, 100%) tested negative. In naturally exposed chickens, the TgSAG1bio-BBMA showed a high sensitivity (98.5%; 95% confidence interval, CI: 90.7–99.9%) and specificity (100%; 95% CI: 85.0–100%) relative to a reference standard established using ELISA, IFAT and MAT. Almost all naturally exposed chickens that were positive in bioassay or by PCR tested positive in the TgSAG1bio-BBMA (93.5%; 95% CI: 77.1–98.9%), while all bioassay- or PCR-negative chickens remained negative (100%; 95% CI: 85.0–100%).Conclusions: The TgSAG1bio-BBMA represents a suitable method for the detection of T. gondii infections in chickens with high sensitivity and specificity, which is comparable or even superior to other tests. Since assays based on this methodology allow for the simultaneous analysis of a single biological sample with respect to multiple analytes, the described assay may represent a component in future multiplex assays for broad serological monitoring of poultry and other farm animals for various pathogens.

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Development and Clinical Evaluation of a Rapid Serodiagnostic Test for Toxoplasmosis of Cats Using Recombinant SAG1 Antigen

Cited by 22 publications

References 23 publications

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

Toxoplasma gondii Infection in Alpine Red Deer (Cervus elaphus): Its Spread and Effects on Fertility

Fluorescent bead-based serological detection of Toxoplasma gondii infection in chickens

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