Optimization and validation of an ELISA using recombinant Toxoplasma gondii matrix antigen 1 for serodiagnosis of the infection

2015 ◽  
Vol 15 (2) ◽  
pp. 56-60 ◽  
Author(s):  
Buyannemekh Tumurjav ◽  
Mohamad Alaa Terkawi ◽  
Houshuang Zhang ◽  
Guohong Zhang ◽  
Honglin Jia ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tissue Cultures ◽  
Chain Reaction ◽  
Serological Tests ◽  
Recombinant Antigens ◽  
Crude Antigen ◽  
Polymerase Chain ◽  
Alternative Sources ◽  
Toxoplasma Gondii Infection

Toxoplasma gondii infection can be diagnosed directly by polymerase chain reaction (PCR), hybridization and isolation of parasites and indirectly with serological methods[4; 5; 18].Although all these tests have shortcomings, serological tests, particularly the enzyme-linked immunosorbent assay (ELISA), seem to be the most practical and economical. The crude antigen prepared from tachyzoites has been traditionally utilized for commercially serological detection kits. However the use of recombinant antigens can be alternative sources of antigens allowing better standardization of the tests and reducing the costs of production requires mass production of the parasite either from the peritoneal fluids of infected mice or from tissue cultures. In spite of the potential advantages of using recombinant antigens in serology tests, their sensitivities have not yet achieved perfect result; therefore, further research on new antigensis extremely desirable [10; 16; 17; 3]. In this context, the Toxoplasma gondii matrix antigen 1 (TgMAG1) known as 65-kDa protein abundantly expressed within the cyst and in the cyst wall surrounding the bradyzoites [15], has documented to be immunogenic during the infection with T. gondii in mouse model and promising reagent for serodiagnosis of toxoplasmosis in humans [15;12; 6]. However, its usefulness has not yet been confirmed in animal toxoplasmosis.In this study, the optimization and validation of E.coli-expressed rTgMAG1as ELISA antigen were describedMongolian Journal of Agricultural Sciences Vol.15(2) 2015; 56-60

scholarly journals Seroprevalence of brucellosis in patients with prolonged fever in Bangladesh

2016 ◽  
Vol 10 (09) ◽  
pp. 939-946 ◽  
Author(s):  
AKM Anisur Rahman ◽  
Dirk Berkvens ◽  
Claude Saegerman ◽  
David Fretin ◽  
Noor Muhammad ◽  
...  
Keyword(s):  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Brucellosis ◽  
Chain Reaction ◽  
Serological Tests ◽  
Human Sera ◽  
Polymerase Chain ◽  
Species Specific ◽  
First Time ◽  
Sera Samples

Introduction: This study describes the seroprevalence of human brucellosis among pyretic patients and detection of Brucella abortus DNA from seropositive pyretic patients using real-time polymerase chain reaction (rtPCR) for the first time in Bangladesh. Methodology: Blood samples were collected from 300 pyretic patients from October 2007 to May 2008 and subjected to three serological tests: Rose-Bengal plate test (RBT), standard tube agglutination test (STAT), and indirect enzyme-linked immunosorbent assay (iELISA). Risk factors were identified by multivariate Firth’s logistic regression analysis. Brucella genus (BCSP31) and species-specific (IS711) rtPCR were applied to six human sera samples. Results: The seroprevalence of brucellosis among pyretic patients was estimated to be 2.0% (95% confidence interval [CI]: 0.74–4.30). The odds of brucellosis seropositivity were 8.9 (95% CI: 1.26–63.0) times higher in pyretic patients who handled goats than those who handled only cattle, whereas the odds of brucellosis seropositivity were 9.7 (95% CI: 1.28–73.68) times higher in pyretic patients who had backache compared to those without backache. B. abortus DNA was amplified from all six human sera that tested positive by RBT, STAT, and iELISA. As the agreement between the tests was very strong, RBT is recommended as a screening test for the diagnosis of human brucellosis in Bangladesh because it is easier to use, cheaper, and faster. Conclusions: Brucellosis among pyretic patients is common, and B. abortus is responsible for brucellosis in such patients. Pyretic patients who handle goats and those with backaches should be screened for brucellosis.

scholarly journals Serodiagnosis of Recently Acquired Toxoplasma gondii Infection Using an Enzyme-Linked Immunosorbent Assay with a Combination of Recombinant Antigens

2000 ◽  
Vol 7 (5) ◽  
pp. 781-787 ◽  
Author(s):  
Shuli Li ◽  
Gina Galvan ◽  
Fausto G. Araujo ◽  
Yasuhiro Suzuki ◽  
Jack S. Remington ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Pregnant Women ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recombinant Antigens ◽  
Single Antigen ◽  
Toxoplasma Gondii Infection

ABSTRACT An enzyme-linked immunosorbent assay (ELISA) using four recombinant antigens of Toxoplasma gondii (rP22, rP25, rP29, and rP35) was used in an attempt to differentiate pregnant women with toxoplasma serologic profiles (TSPs) indicative of recently acquired infections (acute profile) from those with TSPs indicative of infections acquired in the distant past (chronic profile). In general, immunoglobulin G antibodies in sera from women with the acute profile reacted more strongly with the recombinant antigens than did those in sera from women with the chronic profile. However, reactivities differed significantly between antigens that reacted with a single serum and between sera that reacted with a single antigen. Because of these variations, we employed a combination of the four antigens in an ELISA (Comb-ELISA) and evaluated its ability to distinguish pregnant women with the acute profile from those with the chronic profile. Eighteen of 20 (90%) sera from acute-profile women were positive in the Comb-ELISA, whereas 69 of 70 (98.6%) sera from the chronic-profile women were negative. Thus, the Comb-ELISA may be useful for diagnosis of toxoplasmosis in pregnant women and for differentiation between recently acquired infections and infections acquired in the more distant past.

scholarly journals Molecular and serological prevalence of Toxoplasma gondii and Anaplasma spp. infection in goats from Chongqing Municipality, China

Parasite ◽  
2018 ◽  
Vol 25 ◽  
pp. 20 ◽  
Author(s):  
Zuoyong Zhou ◽  
Yutong Wu ◽  
Yiwang Chen ◽  
Zhiying Wang ◽  
Shijun Hu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxoplasma Gondii ◽  
Infection Rate ◽  
Anaplasma Phagocytophilum ◽  
Zoonotic Diseases ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Blood Samples ◽  
Chongqing Municipality ◽  
Polymerase Chain

Toxoplasmosis and anaplasmosis are severe zoonotic diseases, the former caused by Toxoplasma gondii and the latter by Anaplasma spp. In the present study, 332 goat blood samples were randomly collected from Chongqing Municipality, China to screen for T. gondii and Anaplasma spp. We used a polymerase chain reaction (PCR) to detect DNA, and enzyme-linked immunosorbent assay (ELISA) to test for T. gondii antibodies. The prevalence of T. gondii and Anaplasma spp. was 38% and 35% respectively by PCR, and 42% for T. gondii antibodies by ELISA. The co-infection rate by T. gondii and Anaplasma was 13%, where the two predominant pathogens co-infecting were Anaplasma phagocytophilum + A. bovis (10%), followed by T. gondii + A. phagocytophilum (9.64%). While co-infection by three pathogens varied ranging from 1.81% to 5.72%, less than 1% of goats were found to be positive for four pathogens. This is the first investigation of T. gondii and Anaplasma spp. infection in goats from Chongqing.

scholarly journals Prevalence and prevention of brucellosis in cattle in Lebanon

2020 ◽  
Vol 13 (2) ◽  
pp. 364-371 ◽  
Author(s):  
Hussein Hassan ◽  
Ali Salami ◽  
Nada Nehme ◽  
Raed Al Hakeem ◽  
Jeanne El Hage ◽  
...  
Keyword(s):  
Competitive Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Serological Tests ◽  
The Public ◽  
Underdeveloped Countries ◽  
Milk Samples ◽  
Polymerase Chain ◽  
Region 10 ◽  
Insight Into

Background and Aim: Brucellosis is a zoonotic disease caused by the bacterium of the genus Brucella. This disease is present worldwide, especially in developing and underdeveloped countries, where it is endemic. This first-of-its-kind study in Lebanon aimed to assess the prevalence of brucellosis across the country and to determine the efficacy of a vaccine for reducing losses in herds so that its toll on public health is reduced. Materials and Methods: Three hundred and fifty-three blood serum and 261 milk samples were obtained from cows in different areas of Lebanon. The samples were analyzed using serological tests (rose Bengal, milk ring, and indirect enzyme-linked immunosorbent assay [ELISA]) and confirmed with competitive ELISA and polymerase chain reaction. Results: The highest rate of Brucellae was found in the Bekaa region (10%). After vaccination of 5 cows and 13 heifers at different times, the results showed that all the vaccinated animals have developed an immune response to brucellosis 60 days after vaccination. This vaccine can be considered as stable and preventative to protect against brucellosis in animals and thus protect the public from this infection. Conclusion: These findings will provide further insight into designing future targeted awareness interventions and adapted policies as efforts toward reducing the prevalence and prevention of brucellosis in cattle in Lebanon.

scholarly journals Evaluation of a Recombinant Multiepitope Peptide for Serodiagnosis of Toxoplasma gondii Infection

2012 ◽  
Vol 19 (3) ◽  
pp. 338-342 ◽  
Author(s):  
Jianfang Dai ◽  
Min Jiang ◽  
Yanyun Wang ◽  
Lili Qu ◽  
Rujun Gong ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Toxoplasma Gondii ◽  
Diagnostic Tests ◽  
Routine Screening ◽  
Serological Tests ◽  
Recombinant Antigens ◽  
Content Type ◽  
Prevention And Treatment ◽  
Immunosorbent Assays ◽  
Toxoplasma Gondii Infection

ABSTRACTDetection ofToxoplasma gondiiinfection with sensitive and specific methods is a key step in the prevention and treatment of toxoplasmosis. Among the available diagnostic tests, serology is commonly used. Although serological tests give satisfactory results, the production of reliable reagents remains laborious and expensive. There is therefore a real need to acquire specific and effective recombinant antigens for the serodiagnosis ofT. gondiiinfection. In this study, a multiepitope peptide was designed and successfully expressed inEscherichia coli, and then IgG and IgM enzyme-linked immunosorbent assays (ELISAs) were developed and evaluated. Our results showed that the new multiepitope antigen is one of the most promising recombinant antigens which could be used in routine screening of human toxoplasmosis.

scholarly journals 4. Detection of Toxoplasma gondii infection by Polymerase Chain Reaction (PCR) and Histological Examination on Balb/c Mice

2016 ◽  
Vol 1 (2) ◽  
pp. 20-26
Author(s):  
Muhammad Hanafiah ◽  
Dwinna Aliza ◽  
Erdiansyah Rahmi ◽  
Wisnu Nurcahyo
Keyword(s):  
Toxoplasma Gondii ◽  
Histological Examination ◽  
Mononuclear Cells ◽  
Chain Reaction ◽  
Internal Organs ◽  
Tissue Samples ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Toxoplasma Gondii Infection ◽  
The Brain

The purpose of this research was to compare the use of PCR method and histological examination to diagnose toxoplasmosis in tissues of Balb/c mice infected with sporulated oocysts through drinking water. A total of 20 male Balb/c mice aged approximately 2 months were used in this experiment. Each mouse was infected with 1x103 Toxoplasma gondii tachyzoites intraperitoneally. Tissue samples (liver, lung, heart, kidney, and brain) were collected from 5 mice on day 1, day 5, day 7, and day 9 after infection. Samples were then examined by PCR and histological methods. The data collected were analyzed descriptively. The results showed that PCR method was more sensitive than histological examination. PCR examination using primer invitrogen gen can amplify DNA T. gondii at 436 bp of the samples from liver, lung, heart and brain on Day 7 and Day 9 after infection. The histological examination showed that the cyst of toxoplasma was found in the brain while mononuclear cells infiltration was found in other internal organs.

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