scholarly journals Detection of Human Toxoplasma-Specific Immunoglobulins A, M, and G with a Recombinant Toxoplasma gondii Rop2 Protein

1998 ◽  
Vol 5 (5) ◽  
pp. 627-631 ◽  
Author(s):  
Valentina Martin ◽  
Miriam Arcavi ◽  
Graciela Santillan ◽  
Maria Regina R. Amendoeira ◽  
Elizabeth De Souza Neves ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Early Stage ◽  
Humoral Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Systems ◽  
Igg Antibody ◽  
Serological Tests ◽  
Antibody Reactivity ◽  
Group A ◽  
Group D

ABSTRACT The Toxoplasma gondii rhoptry protein Rop2 was expressed in Escherichia coli as a fusion protein containing 44 kDa of the 55-kDa mature Rop2, supplied with six histidyl residues at the N-terminal end (Rop2196–561). Humoral response during Toxoplasma infection of humans was analyzed by immunoglobulin G (IgG), IgA, and IgM enzyme-linked immunosorbent assay with Rop2196–561 as the antigen substrate. The analyzed sera were divided according to T. gondii-specific serological tests (IgG, IgA, or IgM indirect immunofluorescence and IgA or IgM immunosorbent agglutination assay) as group A (IgG+ IgA− IgM−;n = 35), group B (IgG+ IgA+IgM+; n = 21), group C (IgG+IgA+ IgM−; n = 5), and group D (IgG+ IgA− IgM+;n = 16). Twenty-six T. gondii-seronegative sera from individuals with other infections were also included (group E). Anti-Rop2 IgG antibodies were detected in 82.8% of group A sera and in 97.6% of the sera with acute-phase marker immunoglobulins (groups B, C, and D). The percentage of IgA antibody reactivity against Rop2196–561 was 17.1% in group A, 50% in group D, and 80.8% in groups B and C. The percentage of IgM antibody reactivity was 0% in groups A and C and 62% in groups B and D. Sera from group E failed to show IgA, IgM, or IgG antibody reactivity. Since T. gondii Rop2 elicits a strong humoral response from an early stage of infection, it is suggested that recombinant Rop2196–561 would be suitable for use in diagnostic systems, in combination with other T. gondii antigens, to detect specific IgG, IgA, and IgM antibodies.

scholarly journals Detection of Toxoplasma gondii Infections using Virus-Like Particles Displaying T. gondii ROP4 Antigen

2021 ◽  
Vol 59 (6) ◽  
pp. 565-572
Author(s):  
Min-Ju Kim ◽  
Jie Mao ◽  
Hae-Ji Kang ◽  
Ki-Back Chu ◽  
Fu-Shi Quan
Keyword(s):  
Toxoplasma Gondii ◽  
Early Stage ◽  
Dependent Manner ◽  
Igg Antibody ◽  
Serological Tests ◽  
Virus Like Particles ◽  
Highly Sensitive ◽  
Infection Diagnostics ◽  
Tissue Lysate ◽  
Specific Igg

Toxoplasma gondii ME49 infections are typically diagnosed by serological tests. However, serological diagnosis of RH strain-induced toxoplasmosis remains unknown. In order to develop seradiagnosis of above 2 kinds of infections, we generated recombinant virus-like particles (VLPs) displaying the T. gondii rhoptry protein 4 (ROP4) and evaluated their potential in T. gondii ME49 or RH strain infection diagnostics. Mice were orally infected with either the tachyzoites of T. gondii (RH) or cysts of T. gondii (ME49) at various dosages, and sera were collected at regular intervals. ELISA-based serological tests were performed to assess IgG, IgM, and IgA antibody responses against ROP4 VLP antigen and tissue lysate antigen (TLA). Compared to TLA, IgG, IgM, and IgA levels to ROP4 VLP antigen were significantly higher in the sera of T. gondii RH-infected mice 1 and 2 week post-infection (PI). T. gondii-specific IgG antibody was detected at 1, 2, 4, and 8 week PI in the T. gondii ME49-infected mice with infection dose-dependent manner. These results indicated that the ROP4 VLP antigen was highly sensitive antigens detecting T. gondii RH and ME49 antibodies at an early stage.

scholarly journals Impact of Stopping Trastuzumab in Early Breast Cancer: A Population-Based Study in Ontario, Canada

2020 ◽  
Vol 112 (12) ◽  
pp. 1222-1230 ◽  
Author(s):  
Moira Rushton ◽  
Isac Lima ◽  
Meltem Tuna ◽  
Chris Johnson ◽  
Josee Ivars ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cardiac Imaging ◽  
Early Stage ◽  
Population Based ◽  
Left Ventricular Ejection ◽  
Cancer Outcomes ◽  
Adjuvant Trastuzumab ◽  
Group A ◽  
Group B ◽  
Group D

Abstract Background Adjuvant trastuzumab for early-stage (I-III) HER2-positive breast cancer (BC) has led to statistically significant improvement in cancer outcomes but carries a risk of cardiotoxicity. Trastuzumab is discontinued early in many patients for asymptomatic changes in left ventricular ejection fraction. We evaluated the impact of early discontinuation of trastuzumab on cancer outcomes. Methods We conducted a retrospective population-based cohort study of early BC patients treated with adjuvant trastuzumab in Ontario, Canada, 2007-2016. Four groups were analyzed: group A was full treatment, 17-18 cycles trastuzumab; group B was cardiac event (CE) within treatment period; group C was ≤16 cycles, no CEs, stopped within 30 days from last cardiac imaging; and group D was ≤16 cycles, no CEs, stopped more than 30 days from cardiac imaging. Primary outcome was disease-free survival (DFS); secondary outcomes were: overall survival, cancer-specific mortality, and cardiovascular mortality. Sensitivity analyses were performed 14 months after cycle 1 trastuzumab to control for early relapse. Results A total of 5547 patients met the inclusion criteria: group A = 3921, group B = 309, group C = 362, and group D = 955. The 5-year DFS was 94.1% in group A, 80.1% in group B, 81.4% in group C, and 82.4% in group D. Using a Cox model, the hazard ratio for 5-year DFS was 3.15 (95% confidence interval [CI] = 2.13 to 4.65) for group B, 1.94 (95% CI = 1.30 to 2.89) for group C, and 1.92 (95% CI = 1.46 to 2.53) for group D. Overall, 26 patients (0.5%) died of cardiac causes. Conclusions BC patients in Ontario who did not complete adjuvant trastuzumab had a statistically significantly higher risk of BC relapse and death and low incidence of cardiac death. These findings support 1 year of adjuvant trastuzumab in early-stage BC.

Frequency of Toxoplasma gondii in Pork Meat in Ocotlán, Jalisco, Mexico

2010 ◽  
Vol 73 (6) ◽  
pp. 1121-1123 ◽  
Author(s):  
M. L. GALVÁN-RAMIREZ ◽  
A. L. MADRIZ ELISONDO ◽  
C. P. RICO TORRES ◽  
H. LUNA-PASTÉN ◽  
L. R. RODRÍGUEZ PÉREZ ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Pcr Amplification ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Consumption ◽  
Pork Meat ◽  
Igg Antibody ◽  
High Prevalence ◽  
B1 Gene ◽  
Hematoxylin And Eosin ◽  
Antibody Kinetics

Toxoplasmosis is an infection caused by Toxoplasma gondii, an intracellular obligate parasite. Its transmission has usually been attributed to ingestion of undercooked or raw meat. The frequency of T. gondii in pork, the most common meat for human consumption in Jalisco, Mexico, is unknown; in Guadalajara city high prevalence of human toxoplasmosis has been documented. Forty-eight samples of pork meat from butcher shops in Ocotlán city were analyzed. Through bioassay, 50 g of tissue was homogenized in an acidic pepsin solution and inoculated subcutaneously to previously immunosuppressed mice. Blood samples from the mice tail vein were obtained before inoculation and 7, 14, 28, and 45 days postinoculation to analyze anti-Toxoplasma immunoglobulin (Ig) M and IgG antibody kinetics by indirect enzyme-linked immunosorbent assay. For histopathology, small fragments of the brain, lungs, heart, and skeletal muscle were extracted on day 45 and were stained with hematoxylin and eosin. Also, DNA was extracted from the pork meat for PCR amplification of the B1 gene. Even though all pork samples were negative by histopathology and PCR, IgG and IgM antibodies against T. gondii were detected in 1 of the 48 inoculated mice, reflecting a frequency of 2.1% positive pork meat, which is lower than expected but similar to that found in other regions.

scholarly journals Serological Diagnosis of Brucella Infections in Odontocetes

2009 ◽  
Vol 16 (6) ◽  
pp. 906-915 ◽  
Author(s):  
Gabriela Hernández-Mora ◽  
Charles A. Manire ◽  
Rocío González-Barrientos ◽  
Elías Barquero-Calvo ◽  
Caterina Guzmán-Verri ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Brucella Melitensis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Serum Samples ◽  
Igg Antibody ◽  
Serological Tests ◽  
Diagnostic Potential ◽  
Control Serum ◽  
History Of

ABSTRACT Brucella ceti causes disease in Odontoceti. The absence of control serum collections and the diversity of cetaceans have hampered the standardization of serological tests for the diagnosis of cetacean brucellosis. Without a “gold” standard for sensitivity and specificity determination, an alternative approach was followed. We designed an indirect enzyme-linked immunosorbent assay (iELISA) that recognizes immunoglobulins G (IgGs) from 17 odontocete species as a single group. For the standardization, we used Brucella melitensis and Brucella abortus lipopolysaccharides, serum samples from seven resident odontocetes with no history of infectious disease displaying negative rose bengal test (RBT) reactions, and serum samples from seven dolphins infected with B. ceti. We compared the performance of the iELISA with those of the protein G ELISA (gELISA), the competitive ELISA (cELISA), and the immunofluorescence (IF) and dot blot (DB) tests, using 179 odontocete serum samples and RBT as the reference. The diagnostic potential based on sensitivity and specificity of the iELISA was superior to that of gELISA and cELISA. The correlation and agreement between the iELISA and the gELISA were relatively good (R i/g 2 = 0.65 and κi/g = 0.66, respectively), while the correlation and agreement of these two ELISAs with cELISA were low (R i/c 2 = 0.46, R g/c 2 = 0.37 and κi/c = 0.62, κg/c = 0.42). In spite of using the same anti-odontocete IgG antibody, the iELISA was more specific than were the IF and DB tests. An association between high antibody titers and the presence of neurological symptoms in dolphins was observed. The prediction is that iELISA based on broadly cross-reacting anti-dolphin IgG antibody would be a reliable test for the diagnosis of brucellosis in odontocetes, including families not covered in this study.

scholarly journals Reliability of serological tests for COVID-19: Comparison of three immunochromatography test kits for SARS-CoV-2 antibodies

2020 ◽  
Author(s):  
Hidetsugu Fujigaki ◽  
Masao Takemura ◽  
Michiko Osawa ◽  
Aki Sakurai ◽  
Kentaro Nakamoto ◽  
...  
Keyword(s):  
Early Stage ◽  
Serological Test ◽  
Entire Period ◽  
Serum Samples ◽  
Igg Antibody ◽  
Serological Tests ◽  
Igm Antibody ◽  
Potential Clinical Utility ◽  
Almost All ◽  
Positive Results

AbstractBackgroundSeveral immunochromatographic serological test kits have been developed to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific antibodies, but their relative performance and potential clinical utility is unclear.MethodsThree commercially available serological test kits were evaluated using 99 serum samples collected from 29 patients diagnosed with coronavirus disease 2019 (COVID-19).ResultsThe IgM antibody-positive rates of the three serological test kits for samples taken at the early stage of the disease (0–6 days after onset) were 19.0%, 23.8%, and 19.0%, respectively. The IgM antibody-positive rates over the entire period were 21.2%, 60.6%, and 15.2%, respectively. The IgG antibody-positive rates for samples taken after 13 days of onset were 100.0%, 97.6%, and 97.6%, respectively.ConclusionThere were large differences among the results of the three test kits. Only few cases showed positive results for IgM in the early stage of disease and the IgM antibody-positive rates over the entire period were low, suggesting that the kits used in this study were unsuitable for diagnosis of COVID-19. The IgG antibody was positive in almost all samples after 13 days of onset, suggesting that it may be useful for determining infections in the recent past.

scholarly journals SIGNIFICANCE OF SEROLOGICAL MONITORING FOR POULTRY ORNITOBACTERIOSIS THERAPY

2021 ◽  
Vol 22 (1) ◽  
pp. 30-37
Author(s):  
I. K. Avdosieva ◽  
O. I. Chajkovska ◽  
O. B. Basarab ◽  
V. V. Regenchuk
Keyword(s):  
Egg Production ◽  
Early Stage ◽  
Age Groups ◽  
Clinical Signs ◽  
Live Weight ◽  
Suppressive Effect ◽  
Field Strain ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Tests ◽  
Tissue Samples

One of the main problems of poultry diseases is respiratory diseases. Among them a special place is occupied by ornithobacteriosis (ORT). Losses from ORT consist of: direct losses as a result of the disease - death of chickens, increased culling due to lameness, low live weight gain (up to 40%), reduction of carcass categories, reduction of egg production by 6-20%; indirect losses associated with the immune-suppressive effect of ornithobacteria, which increase the risk of other infections and prevent the formation of post-vaccination immunity. The diagnosis is established on the basis of epizootological data, clinical signs, pathological and anatomical changes, bacteriological and serological tests, positive bioassay. In most cases, infections caused by ornithobacteria are not diagnosed in time, the pathogen is difficult to isolate due to complications of other pathogens, or because experts are currently insufficiently aware of the ability of ORT to cause disease. ORT can be isolated by bacteriological method only at an early stage of the disease. The most relevant method of diagnosis is PCR. The advantage of the method is not only the isolation of DNA of individual cells of the pathogen in the sample, but also the ability to detect all serotypes. In addition, PCR is a successful diagnosis in the detection of ORT nucleic acid not only in tissue samples, but also in feces, eggs, dust, which is important for timely diagnosis. Enzyme-linked immunosorbent assay (ELISA) is used to control the presence of ornithobacteriosis in bird. The presence of antibodies to this pathogen in poultry of many species indicates its wide circulation. Thus, when conducting serological monitoring of blood serum from different age groups of broilers aged 1-44 days, the percentage of positive samples ranged from 40 to 100, which indicates the circulation of the field strain of the pathogen ornithobacteriosis. The percentage of positive serum from broilers to ORT was: from 1 to 5 days - from 88 to 50, from 6 to 10 days serum were negative, while at 17, 21 days and from 32 days to the end of cultivation (44 days) – 100 %. The percentage of positive batches at the end of fattening ranged from 42-53 days in the range from 75 to 100%, indicating the circulation of the field strain of the ornithobacteriosis pathogen antibiotic therapy against this disease.

scholarly journals Quantitative SARS-CoV-2 antibody screening of healthcare workers in the southern part of Kyoto city during the COVID-19 peri-pandemic period

2020 ◽  
Author(s):  
Kohei Fujita ◽  
Shinpei Kada ◽  
Osamu Kanai ◽  
Hiroaki Hata ◽  
Takao Odagaki ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Early Stage ◽  
Case Fatality ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igg Antibody ◽  
Kyoto City ◽  
History Of ◽  
Heavy Burden ◽  
Antibody Levels

Background: The coronavirus disease-2019 (COVID-19) pandemic is associated with a heavy burden on the mental and physical health of patients, regional healthcare resources, and global economic activity. While our understanding of the incidence and case-fatality rates increases, data on seroprevalence of antibodies against the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) in healthcare workers during the peri-pandemic period is insufficient. This study quantitatively evaluated seroprevalence of SARS-CoV-2 antibody in healthcare workers in the southern part of Kyoto city, Japan. Methods: We prospectively recruited healthcare workers from a single hospital between April 10 and April 20, 2020. We collected serum samples from these participants and quantitatively evaluated SARS-CoV-2 IgG antibody levels by enzyme-linked immunosorbent assay. Results: Five (5.4%), 15 (16.3%), and 72 (78.3%) participants showed positive, borderline, and negative serum SARS-CoV-2 IgG antibody status, respectively. We found the mean titer associated with each antibody status (overall, positive, borderline, and negative) was clearly differentiated. Participants working at the otolaryngology department and/or having a history of seasonal common cold symptoms had a significantly higher titer of SARS-CoV-2 IgG antibody (p=0.046, p=0.046, respectively). Conclusions: Five (5.4%) and 15 (16.3%) participants tested positive and borderline, respectively, for SARS-CoV-2 IgG antibody during the COVID-19 peri-pandemic period. These rates were higher than expected based on government situation reports. The present findings suggest that COVID-19 was already spread in the southern part of Kyoto city at the early stage of pandemic.

scholarly journals The distribution pattern and growth factor level in platelet-rich fibrin incorporated skin-derived mesenchymal stem cells: An in vitro study

2020 ◽  
Vol 13 (10) ◽  
pp. 2097-2103
Author(s):  
Igo Syaiful Ihsan ◽  
Deya Karsari ◽  
Nora Ertanti ◽  
Aristika Dinaryanti ◽  
Alexander Patera Nugraha ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Growth Factor ◽  
Distribution Pattern ◽  
Enzyme Linked Immunosorbent Assay ◽  
Platelet Rich Fibrin ◽  
Group A ◽  
Group D

Background and Aim: A skin wound in an animal must be cared for to prevent further health issues. Platelet-rich fibrin (PRF) and skin-derived mesenchymal stem cells (SMSCs) have been reported to have potential in increasing the rate of wound healing. This study aimed to analyze the distribution patterns and levels of platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), and transforming growth factor-β (TGF-β) in PRF incorporated with SMSCs. Materials and Methods: This study employed a true experiment (in vitro) design with post-test only performed in the control group alone. PRF and SMSCs were extracted from the blood and skin of 16 rabbits. SMSCs were characterized using immunocytochemistry to examine clusters of differentiation for 45, 73, 90, and 105. PRF was incorporated into the SMSCs and then divided into four groups (N=32/n=8): Group A (PRF only), Group B (PRF+SMSCs, incubated for 1 day), Group C (PRF+SMSCs, incubated for 3 days), and Group D (PRF+SMSCs, incubated for 5 days). Scanning electron microscopy was used to examine the distribution pattern of SMSCs between groups. The supernatant serum (Group A) and supernatant medium culture (Group D) were collected for the measurement of PDGF, IGF, VEGF, and TGF-β using an enzyme-linked immunosorbent assay sandwich kit. An unpaired t-test was conducted to analyze the differences between Groups A and D (p<0.01). Results: Group D had the most morphologically visible SMSCs attached to the PRF, with elongated and pseudopodia cells. There was a significant difference between the levels of growth factor in Groups A and D (p=0.0001; p<0.01). Conclusion: SMSCs were able to adhere to and distribute evenly on the surface of PRF after 5 days of incubation. The PRF incorporated SMSCs contained high levels of PDGF, IGF, VEGF, and TGF- β, which may prove to have potential in enhancing wound healing.

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 Melatonin protects uterus and oviduct exposed to nicotine in mice

2014 ◽  
Vol 7 (1) ◽  
pp. 41-46 ◽  
Author(s):  
Seyedeh Nazanin Seyed Saadat ◽  
Fahimeh Mohammadghasemi ◽  
Sina Khajeh Jahromi ◽  
Mohammad Amin Homafar ◽  
Mostafa Haghiri
Keyword(s):  
Saline Group ◽  
Estradiol Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protective Effects ◽  
Serum Estradiol Level ◽  
Group A ◽  
Group B ◽  
Er Alpha ◽  
Group D ◽  
Estrus Phase

Abstract Smoking is associated with higher infertility risk. The aim of this study was to evaluate protective effects of melatonin on the uterus and oviduct in mice exposed to nicotine. Adult female mice (n=32) were divided into four groups. Group A: control animals received normal saline, Group B: injected with nicotine 40 μg/kg, Group C: injected with melatonin 10 μg, Group D: injected with nicotine 40 μg/kg and melatonin 10 μg. All animals were treated over 15 days intraperitoneally. On the 16th day, animals in the estrus phase were dissected and their uterus and oviducts were removed. Immunohistochemistry was recruited for studying apoptosis and for detection of estrogen receptor (ER) alpha in luminal epithelium of the uterus and oviduct. Enzyme-linked immunosorbent assay was used for serum estradiol level determination. Nicotine in group B decreased estradiol level and ERalpha numbers both in the uterus and oviduct (p<0.05). Co-administration of melatonin-nicotine in Group D ameliorated the histology of the uterus and oviduct, increased ERalpha numbers and reduced apoptosis in the uterus and oviduct compared with the nicotine Group B (p<0.05). This study indicates that nicotine impairs the histology of the uterus and oviduct and co-administration of melatonin-nicotine ameliorates these findings, partly through alteration in ERalpha numbers and reduction of apoptosis

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