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

2020 ◽  
Author(s):  
Benedikt T. Fabian ◽  
Fatima Hedar ◽  
Martin Koethe ◽  
Berit Bangoura ◽  
Pavlo Maksimov ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
High Sensitivity ◽  
Negative Control ◽  
Immunofluorescence Assay ◽  
Farm Animals ◽  
Serological Tests ◽  
Modified Agglutination Test ◽  
Magnetic Capture ◽  
Luminex Technology ◽  
Toxoplasma Gondii Infection

Abstract 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.

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

2020 ◽  
Author(s):  
Benedikt T. Fabian ◽  
Fatima Hedar ◽  
Martin Koethe ◽  
Berit Bangoura ◽  
Pavlo Maksimov ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Small Ruminants ◽  
High Sensitivity ◽  
Negative Control ◽  
Immunofluorescence Assay ◽  
Farm Animals ◽  
Serological Tests ◽  
Modified Agglutination Test ◽  
Magnetic Capture ◽  
Luminex Technology

Abstract Background: Free-ranging chickens are often infected with Toxoplasma gondii. Their infection indicates environmental contamination with T. gondii. The detection of infected birds relies primarily on serological assays. Methods: Here, we established a bead-based multiplex assay (BBMA) using the Luminex technology for the specific and sensitive 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 and bioassay in mice.Results: In experimentally infected chickens, the vast majority (98.5%, n=65/66) of inoculated 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: 90.7-99.9%) and specificity (100%; 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%; 77.1-98.9%), while all bioassay- or PCR-negative chickens remained negative (100%; 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 one component in future multiplex assays for broad serological monitoring of poultry and other farm animals, including pigs or small ruminants, for various pathogens.

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

2020 ◽  
Author(s):  
Benedikt T. Fabian ◽  
Fatima Hedar ◽  
Martin Koethe ◽  
Berit Bangoura ◽  
Pavlo Maksimov ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Small Ruminants ◽  
High Sensitivity ◽  
Negative Control ◽  
Immunofluorescence Assay ◽  
Farm Animals ◽  
Mouse Bioassay ◽  
Serological Tests ◽  
Magnetic Capture ◽  
Luminex Technology

Abstract Background: Especially free-ranging chickens are frequently exposed to Toxoplasma gondii. They are sensitive indicators for environmental contamination with T. gondii oocysts. The detection of infected birds relies primarily on serological assays. Methods: Here, we established a bead-based multiplex assay (BBMA) using the Luminex technology for the specific and sensitive 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. Specific 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 individual 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 and bioassay in mice.Results: In experimentally infected chickens, all chickens were positive by magnetic-capture PCR (100%, n=45/45) and the vast majority (98.5%, n=65/66) of inoculated birds tested seropositive in the BBMA. Most, but not all inoculated and TgSAG1bio-BBMA-positive chickens were also positive in two previously established TgSAG1-ELISAs, an immunofluorescence assay (IFAT) and in a modified agglutination test (MAT). All non-inoculated control animals (n=28) tested negative. In naturally exposed chickens, the TgSAG1bio-BBMA showed a high sensitivity (98.5%; 95% Confidence Interval: 90.7-99.9%) and specificity (100%; 85.0-100%) relative to a reference standard established using results obtained with ELISA, IFAT and MAT. Almost all naturally exposed chickens that were positive in the mouse bioassay or by PCR tested positive in the TgSAG1bio-BBMA (93.5%, 77.1-98.9%), while all mouse bioassay- or PCR-negative chickens remained negative (100%, 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 one component in future multiplex assays for broad serological monitoring of poultry and other farm animals, including pigs or small ruminants, for various pathogens.

scholarly journals Prevalence of Toxoplasma gondii infection in the dairy goat population from an organic farm

2017 ◽  
Vol 73 (11) ◽  
pp. 736-738
Author(s):  
Dawid Jańczak ◽  
Marcin Świątek ◽  
Żaneta Szymańska ◽  
Roman Niżnikowski ◽  
Elżbieta Gołąb
Keyword(s):  
Toxoplasma Gondii ◽  
Public Health Problem ◽  
Farm Animals ◽  
Dairy Goat ◽  
Economic Losses ◽  
Dairy Goats ◽  
Serological Tests ◽  
Organic Farm ◽  
Goat Population ◽  
Toxoplasma Gondii Infection

Protozoal infection of T. gondii is a public health problem and also causes serious economic losses in livestock production in many countries. Farm animals from organic farms are more likely to be infected. The aim of the study was to determine the prevalence of Toxoplasma gondii among 50 dairy goats from an organic farm in the northwestern Poland region and to assess the prevalence of parasite DNA in the milk of infected animals. Serological tests performed by direct agglutination of IgG antibodies against T. gondii were positive in 10% of the tested animals. No parasite DNA was detected in the milk from the seropositive goats. However, the number of tested animals was too small to draw significant epidemiological conclusions.

scholarly journals Toxoplasma gondii infection and toxoplasmosis in farm animals: Risk factors and economic impact

2019 ◽  
Vol 15 ◽  
pp. e00037 ◽  
Author(s):  
S. Stelzer ◽  
W. Basso ◽  
J. Benavides Silván ◽  
L.M. Ortega-Mora ◽  
P. Maksimov ◽  
...  
Keyword(s):  
Risk Factors ◽  
Toxoplasma Gondii ◽  
Economic Impact ◽  
Farm Animals ◽  
Toxoplasma Gondii Infection

Cross-sectional survey of Toxoplasma gondii infection in colony cats from urban Florence (Italy)

2010 ◽  
Vol 12 (4) ◽  
pp. 351-354 ◽  
Author(s):  
Francesca Mancianti ◽  
Simona Nardoni ◽  
Gaetano Ariti ◽  
Dario Parlanti ◽  
Giovanna Giuliani ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Serum Samples ◽  
Cross Sectional Survey ◽  
Nested Polymerase Chain Reaction ◽  
Cross Sectional ◽  
Modified Agglutination Test ◽  
Faecal Samples ◽  
Key Species ◽  
Polymerase Chain ◽  
Toxoplasma Gondii Infection

Cats are the key species in the epidemiology of Toxoplasma gondii infection, even if the proportion of subjects excreting oocysts is low. The aim of the present paper was to obtain information about seroprevalence, oocyst shedding rate and presence of T gondii DNA in faeces collected from an urban population of colony cats in Florence (Tuscany). Fifty European shorthair feral cats were examined for anti- T gondii specific antibodies by a modified agglutination test (MAT), and for oocysts by microscopic examination and for faecal protozoal DNA, by means of a nested polymerase chain reaction (n-PCR) protocol. Twenty-two out of 50 serum samples (44%) were MAT positive. T gondii oocysts were not detected in any of the examined faecal samples. Eight out of 50 faecal specimens (16%) were n-PCR positive and sequencing of the bands was specific for T gondii. Detection by combination of the two methods was higher than single techniques and enhanced the detection of T gondii up to 48%. Our results suggest that the use of MAT plus PCR in faeces may be the best choice for diagnosis of feline toxoplasmosis. Further studies to ascertain the real infectivity of the copro-PCR positive subjects are required.

Seroprevalence and correlates of Toxoplasma gondii infection in dairy cattle in northwest China

2015 ◽  
Vol 60 (4) ◽  
Author(s):  
Qi-Dong Tan ◽  
Xiao-Yu Yang ◽  
Ming-Yang Yin ◽  
Ling-Ying Hu ◽  
Si-Yuan Qin ◽  
...  
Keyword(s):  
Risk Factors ◽  
Toxoplasma Gondii ◽  
Dairy Cattle ◽  
Epidemiological Data ◽  
Northwest China ◽  
Gansu Province ◽  
Serum Samples ◽  
Modified Agglutination Test ◽  
Significant Difference ◽  
Toxoplasma Gondii Infection

AbstractPrevalence of antibodies to Toxoplasma gondii and risk factors with infection were assessed in dairy cattle from Gansu Province and Ningxia Hui Autonomous Region (NXHAR), northwest China. In total, 1657 serum samples were collected and assayed by the modified agglutination test. The overall seroprevalence was 4.83% at a 1:100 cut-off, with titers of 1:100 in 72, 1:200 in 4, 1:400 in 4. Among the risk factors examined, no statistically significant difference was observed between T. gondii seroprevalence and regions or age of dairy cattle in the logistic regression analysis (P>0.05) and left out of the final model. However, numbers of pregnancies of dairy cattle was considered as main risk factor associated with T. gondii infection. Dairy cattle in nulliparity group (8.89%) had 6 times (OR=6.31, 95% CI, 2.69-14.83, P<0.001) higher risk of being seropositive compared to dairy cattle in 3 or above 3 pregnancies group (1.52%), followed by 1 pregnancy group (4.27%) had nearly 3 times (OR=2.89, 95% CI, 1.11-7.52, P = 0.03) higher risk of being seropositive compared to dairy cattle in 3 or above 3 pregnancies group, although no statistical difference was found between 2 pregnancies group and 3 or above 3 pregnancies group (P = 0.70). The results of this survey indicated the presence of T. gondii infection in dairy cattle in Gansu Province and NXHAR, which enriches the epidemiological data of T. gondii infection in dairy cattle in China, and is helpful to strengthen prevention and control of T. gondii infection in dairy cattle in these two regions.

scholarly journals SURVEI SEROLOGIS TOXOPLASMOSIS PADA TERNAK SAPI DI KABUPATEN PRINGSEWU MENGGUNAKAN METODE TOXOPLASMA MODIFIED AGGLUTINATION TEST (To-MAT)

2019 ◽  
Vol 7 (3) ◽  
pp. 270
Author(s):  
Galang B. ◽  
Emantis R. ◽  
Gina Dania P. ◽  
Endah S.
Keyword(s):  
Toxoplasma Gondii ◽  
Agglutination Test ◽  
Test Method ◽  
Cattle Population ◽  
Modified Agglutination Test ◽  
Toxoplasma Gondii Infection

The cattle population in Lampung Province has a relatively high number when compared to several other provinces. Pringsewu is one of the districts in Lampung Province having the supplier potential of beef in Lampung. The higher the population and production, could cause a potential infection of toxoplasmosis to livestock and human. So far the prevalence of toxoplasmosis in Pringsewu District. This study was conducted to determine the level of toxoplasmosis infection in cattle in Pringsewu district. This study used the To-MAT (Toxoplasma Modified Agglutination Test) method to test Toxoplasma gondii infection in blood of cows samples. The result of the study found that at a chronic level there were 23/24 cows positively infected with T.gondii. Whereas in acute level there are 8/24 cows positively infected with T. gondii. In general, the prevalence of toxoplasmosis is 95.8%. Keywords : Toxoplasmosis, Cow, To-MAT (Toxoplasma Modified Agglutination Test) method, Infection

scholarly journals Seroepidemiologi toksoplasmosis pada masyarakat di Desa Kumu Kabupaten Minahasa tahun 2015

2016 ◽  
Vol 4 (1) ◽  
Author(s):  
Vanessa J. T. Seran ◽  
Billy J. Kepel ◽  
Fatimawali .
Keyword(s):  
Toxoplasma Gondii ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Latex Agglutination Test ◽  
Serological Tests ◽  
Cross Sectional ◽  
History Of ◽  
Blood Specimens ◽  
Toxoplasma Gondii Infection ◽  
Specific Symptoms

Abstract: Toxoplasmosis is a disease caused by Toxoplasma gondii in humans and also in animals. Toxoplasma gondii infection is widespread in the world, about 20-90% of the populations have been exposed to this parasite, and most take place without showing specific symptoms. The purpose of this study was to determine the seroepidemiology of toxoplasmosis among the villagers of kumu in district minahasa on 2015. This study was a cross sectional descriptive study. The study population was Kumu Village community. This study using blood specimens to be tested by latex agglutination test and interviews to determine the distribution of toxoplasmosis is based on risk factors. Seropositive obtained if agglutination occurs on the results of serological tests. Respondents consisted of 20 (90.90%) females and 2 (9.10%) men. The results showed 11 (50%) of respondents has toxoplasma seropositivity were distributed by age, sex, level of education, occupation, diet, hygiene, exposure to dogs, exposure to cats, and a history of miscarriage.Keywords: latex agglutination test, parasites, seropositivity, toxoplasma gondiiAbstrak: Toksoplasmosis merupakan penyakit yang disebabkan oleh Toxoplasma gondii pada manusia dan juga pada hewan. Infeksi Toxoplasma gondii tersebar luas di dunia, sekitar 20-90% populasi dunia pernah terpapar parasit ini, dan sebagian besar berlangsung tanpa menunjukkan gejala yang spesifik. Tujuan penelitian ini adalah untuk mengetahui seroepidemiologi toksoplasmosis pada masyarakat di Desa Kumu Kabupaten Minahasa tahun 2015. Penelitian ini merupakan penelitian deskriptif cross sectional. Populasi penelitian ini adalah masyarakat Desa Kumu. Penelitian ini menggunakan spesimen darah untuk diuji dengan uji aglutinasi lateks dan hasil wawancara untuk mengetahui distribusi toksoplasmosis berdasarkan faktor resiko. Seropositif didapat bila terjadi aglutinasi pada hasil tes serologis. Responden terdiri dari 20 (90,90%) wanita dan 2 (9,10%) pria. Hasil penelitian menunjukkan 11 (50%) responden seropositif toksoplasmosis yang terdistribusi berdasarkan umur, jenis kelamin, tingkat pedidikan, pekerjaan, pola makan, higienitas, paparan anjing, paparan kucing, dan riwayat keguguran.Kata kunci: uji aglutinasi lateks, parasit, seropositif, toxoplasma gondii

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