scholarly journals Prevalence of Neospora caninum antibodies in sheep flocks of Uberlândia county, MG

2010 ◽  
Vol 19 (3) ◽  
pp. 148-151 ◽  
Author(s):  
Sandra Renata Sampaio Salaberry ◽  
Liria Hiromi Okuda ◽  
Alessandra Figueiredo de Castro Nassar ◽  
Jacqueline Ribeiro de Castro ◽  
Anna Monteiro Correia Lima-Ribeiro
Keyword(s):  
Risk Factors ◽  
Indirect Immunofluorescence ◽  
Neospora Caninum ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Parasitic Disease ◽  
Blood Samples ◽  
Sheep Blood ◽  
Wild Canids ◽  
Significant Difference

Neosporosis is a parasitic disease that occurs in sheep and is associated with reproductive failure. The aim of this study was to verify seroprevalence of Neospora caninum antibodies in 12 sheep flocks in Uberlândia county, Minas Gerais, Brazil, using Indirect Immunofluorescence Assay (IFA). A total of 334 sheep blood samples were analyzed and an epidemiologic questionnaire was applied for each farm in order to correlate with risk factors of neosporosis: gender, age, breed, abortion problems, contact with cattle and dogs, and presence of wild canids. The prevalence of Neospora caninum antibodies in sheep was 8.1%, with titers ranging from 50 to 3200 and 83.3% of farms detected at least one ovine reactive. There was statistically significant difference in flocks with abortion problems associated with the presence of Neospora caninum antibodies (p < 0.05).

scholarly journals Seroprevalence and risk factors associated with infection by Neospora caninum of dairy cattle in the state of Alagoas, Brazil

2012 ◽  
Vol 32 (10) ◽  
pp. 1009-1013 ◽  
Author(s):  
Maria E. Sousa ◽  
J.N. Porto Wagnner ◽  
Pedro P.F. Albuquerque ◽  
Orestes L. Souza Neto ◽  
Eduardo B. Faria ◽  
...  
Keyword(s):  
Risk Factors ◽  
Dairy Cattle ◽  
Indirect Immunofluorescence ◽  
Neospora Caninum ◽  
Infection Risk ◽  
Control Measures ◽  
Risk Indicators ◽  
Blood Samples ◽  
Factors Associated ◽  
Serological Investigation

The objective of this study was to investigate the prevalence of anti-Neospora caninum antibodies in cattle from milk producing farms of the microregion of Batalha, state of Alagoas, Brazil, as well as to identify the risk factors associated with the infection. Blood samples were collected from 1,004 cattle of 17 farms for the serological investigation regarding the presence of anti-N. caninum antibodies by the Indirect Immunofluorescence Reaction Technique (IMRT). From the total amount of samples analyzed, 77/1,004 (7.67%) were positive and 927/1,004 (92.33%) were negative. The logistical regression identified that cattle from farms without consortium breeding have an infection risk 6.33 (p<0.001; C.I. 2.89-13.10) times higher than cattle from farms with that type of breeding. Cattle from farms where the aborted fetuses are not adequately buried have an infection risk 3.04 (p<0.001; C.I. 1.64-5.63) times higher than cattle from farms with adequate destination of these fetuses. Infection by N. caninum occurs in cattle of the investigated region. The factors identified in our study can be used as risk indicators, so that control measures could be implemented to avoid infection by N. caninum in the herds of this region.

scholarly journals A Novel Engineered Single-Chain Antibody Fragment for Targeting Pediatric Philadelphia Chromosome-like Acute Lymphoblastic Leukemia

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 36-36
Author(s):  
Sara M.A. Mohamed ◽  
Keith Sia ◽  
Karl-Heinz Friedrich ◽  
Andreas Wohlmann ◽  
Savvas Savvides ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Conflicts Of Interest ◽  
Lymphoblastic Leukemia ◽  
Statistical Significance ◽  
Immunofluorescence Assay ◽  
Laser Scanning Microscopy ◽  
Indirect Immunofluorescence Assay ◽  
Single Chain ◽  
Genetic Characteristics ◽  
Significant Difference

Introduction: Philadelphia-like acute lymphoblastic leukaemia (Ph-like ALL) is a high-risk ALL subtype characterized by an inferior survival rate and chemotherapeutic drug resistance (Tasian et al, Blood 130: 2064-2072, 2017). Around 50% of Ph-like ALL cases harbour gene rearrangements leading to the overexpression of cytokine receptor-like factor 2 (CRLF2) (Loh et al, Blood 121: 485-488, 2013). CRLF2 (also known as thymic stromal lymphopoietin receptor, TSLPR) heterodimerizes with the interleukin-7 receptor alpha (IL-7Rα) subunit to form the functional TSLPR. Upon TSLP binding, CRLF2 activates the JAK/STAT signalling pathway, leading to enhanced proliferation and survival of leukemia cells resulting in poor outcomes in patients (Harvey et al, Blood 115: 5312-5321, 2010). The extracellular overexpression of CRLF2 and association with poor prognosis suggest the translational value of designing precision-based therapeutics targeting CRLF2 in Ph-like ALL. Although conventional immunotherapy using full-sized antibodies is a promising treatment strategy that can improve treatment efficiency and minimize off-target toxicity, their clinical translation is challenging due to the high production cost and large size affecting targeting efficiency (Holliger et al, Nat Biotech 23: 1126-1136, 2005). Herein, we validated a novel single-chain variable fragment against CRLF2 (CRLF2-ScFv) for targeting Ph-like ALL cells. Methods: A CRLF2-rearranged Ph-like ALL cell line (MHH-CALL-4) was lentivirally transduced with a CRLF2-targeting shRNA driven by an inducible promoter, enabling the inducible knockdown of CRLF2. CRLF2 expression in MHH-CALL-4 cells after shRNA induction (KD-CALL-4) was evaluated using fluorescence-activated cell sorting (FACS). The cellular association of the CRLF2-ScFv was determined in MHH-CALL-4 and KD-CALL-4 at 4° and 37°C using an indirect immunofluorescence assay. Confocal laser scanning microscopy was used to visualize and compare the cellular association of CRLF2-ScFv in MHH-CALL-4 and KD-CALL-4. The cellular association of CRLF2-ScFv was also investigated ex vivo using a small panel of Ph-like and non-Ph-like ALL patient-derived xenografts (PDXs), representing similar immunophenotype and genetic characteristics to their original disease subtypes (Liem et al, Blood 103: 3905-3914, 2004), and peripheral blood mononuclear cells (PBMCs) to investigate the non-selective association. CRLF2 expression in MHH-CALL-4 and Ph-like ALL PDX cells was quantified using indirect immunofluorescence assay. The downstream impact of CRLF2-ScFv on STAT5 phosphorylation (pSTAT5) was determined by FACS either with or without TSLP stimulation. The statistical significance was tested using Unpaired unequal variances t-test or one-way ANOVA followed by multiple comparisons test. Statistical significance was considered when P ≤ 0.05. All experiments were performed in triplicate. Results: KD-CALL-4 showed a 75% reduction in CRLF2 expression compared with MHH-CALL-4 cells (P = 0.0009). CRLF2-ScFv exhibited a 94% higher association with MHH-CALL-4 compared with KD-CALL-4 cells at 37°C (P = 0.0013). The association of CRLF2-ScFv with MHH-CALL-4 cells was reduced by 75% at the non-proliferating state of cells at 4°C compared to 37°C (P = 0.006). Orthogonally viewed confocal microscopy images showed 82% higher intracellular uptake of CRLF2-ScFv in MHH-CALL-4 compared to KD-CALL-4 cells (P = 0.0003). CRLF2-ScFv showed &gt;80% higher association with a Ph-like PDX sample compared with a control CRLF2low PDX and PBMCs (P &lt; 0.001). Of note, a Ph-like ALL PDX exhibited only one-third of the association with CRLF2-ScFv compared with MHH-CALL-4 cells (P = 0.04), corresponding to the significant difference in CRLF2 surface expression (P = 0.01). CRLF2-ScFv significantly reduced pSTAT5 expression in MHH-CALL-4 cells (P = 0.05) and prevented TSLP-induced STAT5 phosphorylation (P = 0.01), suggesting competition with the TSLP binding site. Conclusion: CRLF2-ScFv is a selective targeting moiety for CRLF2 with a significant internalization potential and receptor antagonistic effect, highlighting the therapeutic implications for targeting Ph-like ALL. Keywords: CRLF2, ScFv, STAT5 phosphorylation, Patient-Derived Xenografts. Disclosures No relevant conflicts of interest to declare.

Evaluation of Epstein-Barr Virus Indirect Immunofluorescence Assay Results

2020 ◽  
Vol 4 (1) ◽  
Author(s):  
Gorgun S ◽  
◽  
Havuz SG ◽  
Mehel D ◽  
◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Epstein Barr Virus ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Barr Virus ◽  
Ebv Infection ◽  
Study Results ◽  
Significant Difference ◽  
Child Patients ◽  
Epstein Barr

Aim: Epstein-Barr Virus (EBV) causes Infectious Mononucleosis (IM) and chronic active EBV infections, malignant diseases such as Burkitt’s lymphoma, nasopharyngeal carcinoma, and posttransplant lymphoproliferative disease. More than 90% of children are infected with EBV until the age of six. The aim of this study was to diagnose EBV infection using Indirect Immunofluorescence Assay (IIFA) method and to evaluate these patients clinically. Methods: The tests were studied by using EBV Indirect Immunofluorescence Assay (IIFA) method. A total of 247 patients, 186 adults, and 61 children, were included in the study. Results: Five (2.7%) of the adults were EBV-Capsid Antigen (EBV-CA) IgM positive, 175 (94.1%) were IgG positive and 6 (3.2%) were seronegative. 10 (16.4%) of the child patients were IgM positive, which is considered as an acute IM infection marker, whereas the child patients had a significantly higher IgM rate than adults (p <0.001). 39 (63.9%) of the child patients were IgG positive and 12 (19.7%) of them were seronegative. The rate of IgG positivity in children was significantly lower than in adults (p <0.001). There was no significant difference between the genders in terms of IgM and IgG positivity rates in both adults and children (p >0.05 for each). Conclusion: These results suggest that the presence of an acute EBV infection should be considered when the patient has viral diseases with similar clinical picture especially for children.

Evaluation of Epstein-Barr Virus Indirect Immunofluorescence Assay Results

2020 ◽  
Author(s):  
Selim Gorgun ◽  
Seda Gudul Havuz ◽  
Dursun M. Mehel
Keyword(s):  
Indirect Immunofluorescence ◽  
Epstein Barr Virus ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Barr Virus ◽  
Ebv Infection ◽  
Significant Difference ◽  
Child Patients ◽  
Epstein Barr ◽  
Acute Infectious Mononucleosis

Abstract Background: The aim of this study was to diagnose Epstein-Barr virüs (EBV) infection using indirect immunofluorescence assay (IIFA) method and to evaluate these patients clinically. More than 90% of children are infected with EBV until the age of six. Methods: The tests were studied by using EBV IIFA method. A total of 247 patients, 186 adults, and 61 children, were included in the study. Results: 5 (2.7%) of the adults were EBV-Capsid antigen (EBV-CA) IgM positive, 175 (94.1%) were IgG positive and 6 (3.2%) were seronegative. 10 (16.4%) of the child patients were IgM positive, which is considered as an acute infectious mononucleosis (IM) infection marker, whereas the child patients had a significantly higher IgM rate than adults (p <0.001). 39 (63.9%) of the child patients were IgG positive and 12 (19.7%) of them were seronegative. The rate of IgG positivity in children was significantly lower than in adults (p <0.001).There was no significant difference between the genders in terms of IgM and IgG positivity rates in both adults and children(p> 0.05 for each).Conclusion: These results suggest that the presence of an acute EBV infection should be considered when the patient has viral diseases with similar clinical picture especially for children.

scholarly journals Risk factors and anti-Toxoplasma gondii and Neospora caninum antibody occurrence in dogs in João Pessoa, Paraíba state, Northeastern Brazil

2018 ◽  
Vol 27 (2) ◽  
pp. 242-247 ◽  
Author(s):  
Arthur Willian de Lima Brasil ◽  
Roberta Nunes Parentoni ◽  
José Givanildo da Silva ◽  
Carolina de Sousa Américo Batista Santos ◽  
Rinaldo Aparecido Mota ◽  
...  
Keyword(s):  
Risk Factors ◽  
Toxoplasma Gondii ◽  
Indirect Immunofluorescence ◽  
Neospora Caninum ◽  
Northeastern Brazil ◽  
Blood Samples ◽  
Two Agents ◽  
Forested Areas ◽  
Caninum Antibody ◽  
The City

Abstract The aim of this study was to determine the occurrence of infections due to Toxoplasma gondii and Neospora caninum and corresponding risk factors among dogs attended at veterinary clinics in the city of João Pessoa, Paraíba, northeastern Brazil. Blood samples were collected from 384 dogs that were attended at 34 veterinary clinics between April 2015 and May 2016. These two agents were diagnosed through the indirect immunofluorescence reaction (IFAT). Among the 384 animals evaluated, 37 (9.6%) were positive for T. gondii, with titers ranging from 16 to 512. Six dogs (1.6%) were positive for N. caninum, with titers of 50 to 200. Access to the streets (OR = 4.60; 95% CI = 1.74-12.20) and environments close to forested areas (OR = 2.79; 95% CI = 1.32-5.93) were found to be risk factors for T. gondii infection. The dogs attended at veterinary clinics in João Pessoa are exposed to infections caused by T. gondii and N. caninum. Dog owners should avoid having access to the street or contact with forest environments.

scholarly journals The antinuclear antibody HEp-2 indirect immunofluorescence assay: a survey of laboratory performance, pattern recognition and interpretation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anne E. Tebo ◽  
Robert L. Schmidt ◽  
Kamran Kadkhoda ◽  
Lisa K. Peterson ◽  
Edward K. L. Chan ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Clinical Laboratory ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Medical Laboratory ◽  
Nuclear Staining ◽  
International Consensus ◽  
In Vitro Diagnostic ◽  
Traditional Pattern

Abstract Background To evaluate the interpretation and reporting of antinuclear antibodies (ANA) by indirect immunofluorescence assay (IFA) using HEp-2 substrates based on common practice and guidance by the International Consensus on ANA patterns (ICAP). Method Participants included two groups [16 clinical laboratories (CL) and 8 in vitro diagnostic manufacturers (IVD)] recruited via an email sent to the Association of Medical Laboratory Immunologists (AMLI) membership. Twelve (n = 12) pre-qualified specimens were distributed to participants for testing, interpretation and reporting HEp-2 IFA. Results obtained were analyzed for accuracy with the intended and consensus response for three main categorical patterns (nuclear, cytoplasmic and mitotic), common patterns and ICAP report nomenclatures. The distributions of antibody titers of specimens were also compared. Results Laboratories differed in the categorical patterns reported; 8 reporting all patterns, 3 reporting only nuclear patterns and 5 reporting nuclear patterns with various combinations of other patterns. For all participants, accuracy with the intended response for the categorical nuclear pattern was excellent at 99% [95% confidence interval (CI): 97–100%] compared to 78% [95% CI 67–88%] for the cytoplasmic, and 93% [95% CI 86%–100%] for mitotic patterns. The accuracy was 13% greater for the common nomenclature [87%, 95% CI 82–90%] compared to the ICAP nomenclature [74%, 95% CI 68–79%] for all participants. Participants reporting all three main categories demonstrated better performances compared to those reporting 2 or less categorical patterns. The average accuracies varied between participant groups, however, with the lowest and most variable performances for cytoplasmic pattern specimens. The reported titers for all specimens varied, with the least variability for nuclear patterns and most titer variability associated with cytoplasmic patterns. Conclusions Our study demonstrated significant accuracy for all participants in identifying the categorical nuclear staining as well as traditional pattern assignments for nuclear patterns. However, there was less consistency in reporting cytoplasmic and mitotic patterns, with implications for assigning competencies and training for clinical laboratory personnel.

scholarly journals Assessment of antinuclear antibodies by indirect immunofluorescence assay: report from a survey by the American Association of Medical Laboratory Immunologists

2020 ◽  
Vol 58 (9) ◽  
pp. 1489-1497 ◽  
Author(s):  
Lisa K. Peterson ◽  
Anne E. Tebo ◽  
Mark H. Wener ◽  
Susan S. Copple ◽  
Marvin J. Fritzler
Keyword(s):  
Indirect Immunofluorescence ◽  
Antinuclear Antibodies ◽  
Current Practice ◽  
Clinical Laboratory ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Medical Laboratory ◽  
International Consensus ◽  
Clinical Laboratories ◽  
Reading Interpretation

AbstractBackgroundThe indirect immunofluorescence assay (IFA) using HEp-2 cell substrates is the preferred method by some for detecting antinuclear antibodies (ANA) as it demonstrates a number of characteristic staining patterns that reflect the cellular components bound as well as semi-quantitative results. Lack of harmonized nomenclature for HEp-2 IFA patterns, subjectivity in interpretation and variability in the number of patterns reported by different laboratories pose significant harmonization challenges. The main objectives of this study were to assess current practice in laboratory assessment of HEp-2 IFA, identify gaps and define strategies to improve reading, interpretation and reporting.MethodsWe developed and administered a 24-item survey based on four domains: educational and professional background of participants, current practice of HEp-2 IFA testing and training, gap assessment and the perceived value of International Consensus on Antinuclear Antibody Patterns (ICAP) and other factors in HEp-2 IFA assessment. The Association of Medical Laboratory Immunologists (AMLI) and American Society for Clinical Pathology administered the survey from April 1 to June 30, 2018, to members involved in ANA testing. This report summarizes the survey results and discussion from a dry workshop held during the 2019 AMLI annual meeting.ResultsOne hundred and seventy-nine (n = 179) responses were obtained where a significant number were clinical laboratory scientists (46%), laboratory directors (24%), supervisors (13%) or others (17%). A majority of respondents agreed on the need to standardize nomenclature and reporting of HEp-2 IFA results. About 55% were aware of the ICAP initiative; however, among those aware, a significant majority thought its guidance on HEp-2 IFA nomenclature and reporting is of value to clinical laboratories. To improve ICAP awareness and further enhance HEp-2 IFA assessment, increased collaboration between ICAP and the clinical laboratory community was suggested with emphasis on education and availability of reference materials.ConclusionsBased on these suggestions, future efforts to optimize HEp-2 IFA reading, interpretation and reporting would benefit from more hands-on training of laboratory personnel as well as continuous collaboration between professional organizations, in vitro diagnostic manufacturers and clinical laboratories.

scholarly journals A case of human monocytic ehrlichiosis in Serbia

2014 ◽  
Vol 142 (1-2) ◽  
pp. 79-82 ◽  
Author(s):  
Bogdan Arsic ◽  
Ana Gligic ◽  
Elizabeta Ristanovic ◽  
Branislav Lako ◽  
Aleksandar Potkonjak ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Clinical Picture ◽  
Peripheral Blood ◽  
Immunofluorescence Assay ◽  
High Fever ◽  
Tick Bite ◽  
Indirect Immunofluorescence Assay ◽  
Ehrlichia Chaffeensis ◽  
Serum Samples ◽  
History Of

Introduction. Ehrlichiosis is a bacterial zoonosis transmitted by hematophagous arthropods - ticks. In humans, it occurs as monocytic, granulocytic, and ewingii ehrlichiosis. Pathological process is based on parasitic presence of Ehrlichia organisms within peripheral blood cells - monocytes and granulocytes. Case Outline. Fifty-two year old patient was admitted to hospital due to high fever of over 40?C that lasted two days, accompanied with chills, muscle aches, malaise, loss of appetite, headache, confusion, breathing difficulties, and mild dry cough. The history suggested tick bite that occurred seven days before the onset of disease. Doxycycline was introduced and administered for 14 days, causing the disease to subside. Indirect immunofluorescence assay was used to analyze three serum samples obtained from this patient for Ehrlichia chaffeensis antibodies, and peripheral blood smear was evaluated for the presence of Ehrlichia and Ehrlichia aggregation into morulae. Conclusion. Ehrlichiosis should be considered in each case where there is a history of tick bite together with the clinical picture (high fever, chills, muscle aches, headache, generalized weakness and malaise, and possible maculopapular rash). The presence of Ehrlichia chaffeensis antibodies was confirmed in a patient with the history of tick bite, appropriate clinical picture and indirect immunofluorescence assay. This confirmed the presence of human monocytotropic ehrlichiosis, a disease that is uncommonly identified in our country.

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