Evaluation of a Rapid Device for Serological Diagnosis of Leishmania infantum Infection in Dogs as an Alternative to Immunofluorescence Assay and Western Blotting

ABSTRACTIn this study, we compared a rapid immunochromatographic test (Speed Leish K; BVT Groupe Virbac, La Seyne sur Mer, France) with an indirect immunofluorescence assay (IFAT) and Western blotting (WB) for the detection ofLeishmania infantumantibodies in dogs. A total of 250 serum samples were collected from 125L. infantum-positive and 125L. infantum-negative dogs. Among the positive samples, 81 were strongly positive at low IFAT dilutions, while 44 were low-reactivity sera (IFAT titers, 1:40 to 1:80). The sensitivity and specificity of the Speed Leish K were 96.3% and 100%, respectively, compared with those of the IFAT. When IFAT low-reactivity sera (titers, 1:40 or 1:80) were tested with the Speed Leish K, using WB results as a reference, the sensitivities were 93.75% for sera with a 1:80 titer and 73.33% for sera with a 1:40 titer, and the specificity was 100%. The Speed Leish K is easy to use and performs well, so it can be considered a quick and reliable tool for the diagnosis ofL. infantuminfection in dogs.

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Comparison of Enzyme-Linked Immunosorbent Assay, Western Blotting, Microagglutination, Indirect Immunofluorescence Assay, and Flow Cytometry for Serological Diagnosis of Tularemia

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.11.6.1008-1015.2004 ◽

2004 ◽

Vol 11 (6) ◽

pp. 1008-1015 ◽

Cited By ~ 65

Author(s):

Mustafa Porsch-Özcürümez ◽

Nele Kischel ◽

Heidi Priebe ◽

Wolf Splettstösser ◽

Ernst-Jürgen Finke ◽

...

Keyword(s):

Flow Cytometry ◽

Western Blotting ◽

Francisella Tularensis ◽

Immunosorbent Assay ◽

Indirect Immunofluorescence ◽

Immunofluorescence Assay ◽

Indirect Immunofluorescence Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Diagnostic Sensitivity And Specificity

ABSTRACT The serodiagnostic efficiencies of five different approaches to detecting antibodies (immunoglobulins G, A, and M) developed in clinically proven infections with Francisella tularensis have been assessed. Fifty serum samples from patients suffering from tularemia during an outbreak in Sweden were compared with samples from 50 healthy blood donors (controls) by using an enzyme-linked immunosorbent assay (ELISA), microagglutination (MA), Western blotting (WB), an indirect immunofluorescence assay (IIFA), and flow cytometry (FC). ELISA, WB, and FC were based on the use of preparations of lipopolysaccharides (LPS) of the live vaccine strain of Francisella tularensis subsp. holarctica (ATCC 29684) as a capture antigen. Whole methanol-fixed bacteria were used for IIFA and MA. Optimized protocols yielded a diagnostic sensitivity and specificity of 100% for WB, MA, and FC, 98% for ELISA, and 93% for IIFA. A total of 6,632 serum samples from individuals between the ages of 18 and 79 years, representatively recruited from all regions of Germany, were screened to estimate and confirm the positive predictive value (PVpos) of the ELISA. Serum samples from 15 (0.226%) individuals tested positive for F. tularensis-specific antibodies by ELISA and confirmatory WB. The resulting prevalence-dependent PVpos of 10.2% and specificity of 98.1% were consistent with our findings for tularemia patients and controls. We conclude that the combined usage of a screening ELISA and a confirmatory WB based on LPS as a common antigen, as well as the MA, is a suitable serodiagnostic tool, while the quality of the IIFA is hampered by subjective variations of the results. FC is a promising new approach that might be improved further in terms of multiplex analyses or high-throughput applications.

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A case of human monocytic ehrlichiosis in Serbia

Srpski arhiv za celokupno lekarstvo ◽

10.2298/sarh1402079a ◽

2014 ◽

Vol 142 (1-2) ◽

pp. 79-82 ◽

Cited By ~ 3

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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Comparison of Coombs Gel Test with ELISA and Standard Tube Agglutination Tests Used in Serological Diagnosis of Brucellosis

Infectious Diseases and Clinical Microbiology ◽

10.36519/idcm.2019.0024 ◽

2020 ◽

Vol 2 (1) ◽

pp. 1-7 ◽

Cited By ~ 1

Author(s):

Cigdem Akalan Kuyumcu ◽

Serpil Erol ◽

Rıza Adaleti ◽

Seniha Senbayrak ◽

Secil Deniz ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Diagnostic Tests ◽

Serological Test ◽

High Sensitivity ◽

Serological Diagnosis ◽

Control Groups ◽

Serum Samples ◽

Serological Tests ◽

Reliable Test ◽

And Control

Objective: Serological tests are the most commonly used tests in the diagnosis of brucellosis; however, each serological test has some drawbacks. In this study, we aimed to determine the value of the Brucella Coombs gel test (BCGT) in the serological diagnosis of brucellosis in comparison with Standard tube agglutination (STA) and ELISA tests. Materials and Methods: The study included 42 patients who were considered to have brucellosis as a preliminary diagnosis. BCGT, Brucella-IgM/IgG ELISA, and STA tests were performed from serum samples of the patients. The correlation of the diagnostic tests was analyzed using Cohen’s Kappa Analysis. Results: Twenty-seven (64.2%) of 42 patients were diagnosed with brucellosis according to their medical history and clinical and serological tests. The sensitivity and specificity of BCGT to diagnose brucellosis was 96.2%, and 100%, respectively. The sensitivity and specificity for the diagnosis of brucellosis 62.9% and 100% for STA, respectively; 33.3% and 66.6% for Brucella-IgM; and 66.6% and 100% for Brucella-IgG. BCGT was significantly correlated with STA (κ= 0.590) and Brucella-IgG (κ=0.539) Conclusion: BCGT can be utilized as a simple and reliable test in the diagnosis of brucellosis with high sensitivity and specificity. Nevertheless, the sensitivity and specificity of BCGT should be demonstrated by comprehensive studies, including culture-confirmed cases and control groups.

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Subtractive Phage Display Selection from Canine Visceral Leishmaniasis Identifies Novel Epitopes That Mimic Leishmania infantum Antigens with Potential Serodiagnosis Applications

Clinical and Vaccine Immunology ◽

10.1128/cvi.00583-13 ◽

2013 ◽

Vol 21 (1) ◽

pp. 96-106 ◽

Cited By ~ 18

Author(s):

Lourena E. Costa ◽

Mayara I. S. Lima ◽

Miguel A. Chávez-Fumagalli ◽

Daniel Menezes-Souza ◽

Vivian T. Martins ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Phage Display ◽

Leishmania Infantum ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Phage Display Technology ◽

Predictive Values ◽

Content Type ◽

Phage Elisa

ABSTRACTVisceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy andTrypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected withLeishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n= 31) compared to those from vaccinated dogs (n= 21), experimentally infected dogs with cross-reactive parasites (n= 23), and healthy controls (n= 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no cross-reactivity withT. cruzi- orEhrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes ofL. infantumantigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programs.

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Serum-Positive and -Negative AQP4 Antibody NMO in Chinese Patients

Canadian Journal of Neurological Sciences / Journal Canadien des Sciences Neurologiques ◽

10.1017/s0317167100013287 ◽

2012 ◽

Vol 39 (2) ◽

pp. 232-235 ◽

Cited By ~ 3

Author(s):

Youming Long ◽

Zhengqi Lu ◽

Xueqiang Hu

Keyword(s):

Indirect Immunofluorescence ◽

Immunofluorescence Assay ◽

Chinese Patients ◽

Igg Antibodies ◽

Serum Samples ◽

Positive Antibody ◽

A Cell ◽

The Central Nervous System ◽

Aqp4 Antibody ◽

Virchow Robin Space

Objective:To compare the clinical features of our sero-negative and sero-positive neuromyelitis optica (NMO) patients.Methods:Thirty-nine patients with NMO were recruited and analyzed retrospectively. Serum aquaporin 4 (AQP4) antibody status was determined by a cell-based assay. For the sero-negative patients, cerebrospinal fluid (CSF) and serum samples were re-tested using the cell-based assay and an indirect immunofluorescence assay.Results:By the cell-based assay, 30 patients (76.92%, 30/39), were positive for AQP4 antibodies in serum and 37 patients (94.9%, 37/39), had a CSF-positive antibody status. Seven NMO patients (17.9%, 7/39) were sero-negative by the cell-based assay but demonstrated positive CSF results. By indirect immunofluorescence, the remaining two patients, who had no AQP4 antibodies in serum or CSF by the cell-based assay, were positive for IgG antibodies in serum, which selectively targeted the central nervous system microvessels, pia, subpia, Virchow-Robin space, kidney, and stomach. There were no significant differences between the sero-positive and sero-negative NMO groups among their demographic and clinical data.Conclusions:Repeating the test using a different assay or CSF is helpful to clarify whether sero-negative NMO patients do in fact carry AQP4 antibodies.

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Evaluation of Four Commercially Available Epstein-Barr Virus Enzyme Immunoassays with an Immunofluorescence Assay as the Reference Method

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.10.1.78-82.2003 ◽

2003 ◽

Vol 10 (1) ◽

pp. 78-82 ◽

Cited By ~ 40

Author(s):

Barbara C. Gärtner ◽

Ralf D. Hess ◽

Dirk Bandt ◽

Alexander Kruse ◽

Axel Rethwilm ◽

...

Keyword(s):

Indirect Immunofluorescence ◽

Epstein Barr Virus ◽

Reference Method ◽

Immunofluorescence Assay ◽

Parameter Analysis ◽

Indirect Immunofluorescence Assay ◽

Serum Samples ◽

Enzyme Immunoassays ◽

Barr Virus ◽

Epstein Barr

ABSTRACT Four commercially available enzyme immunoassays (EIAs) (Novitec, Biotest, Virotech, and DiaSorin) were evaluated, with an indirect immunofluorescence assay as the reference method, for Epstein-Barr virus (EBV) VCA (viral capsid antigen) immunoglobulin G (IgG), VCA IgM, or EBNA (EBV nuclear antigen) IgG at three different locations (Homburg, Stuttgart, and Dresden). Serum samples from 66 immunocompetent patients with infectious mononucleosis, 73 patients without prior EBV infection, and 96 patients with past EBV infections and 29 serum samples with possible cross-reactions to other herpesviruses were included. In addition, 25 samples from an extensively pretested panel that is commercially available (Boston Biomedica) were tested. Each sample was tested at only one location. The four EIAs varied considerably in performance. When analyzing for EBV diagnosis, the Novitec assay performed the best, with 4.9% discrepant diagnoses, followed by the Biotest, Virotech, and DiaSorin assays, with 6.8, 11.7, and 14.0% discrepant diagnoses, respectively. On the basis of single-parameter analysis, the Novitec assay also showed the lowest number of discrepant results, with 3.5%, compared with the Virotech, Biotest, and DiaSorin assays, which produced 5.4, 6.4, and 8.6% discrepant results, respectively. VCA assays using affinity-purified native antigens performed better than assays with recombinant or synthetic antigens. The synthetic EBNA-1s showed the lowest concordance with the reference compared to recombinant p72. Commercially available EBV EIAs differed considerably in performance; however, some proved to be reliable and convenient alternatives to the indirect immunofluorescence assay for routine diagnostics. Native antigens, rather than synthetic peptides, are favored for EBV serology testing.

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Evaluation of a New Immunochromatographic Test Using Recombinant Antigen B8/1 for Diagnosis of Cystic Echinococcosis

Journal of Clinical Microbiology ◽

10.1128/jcm.02157-15 ◽

2015 ◽

Vol 53 (12) ◽

pp. 3859-3863 ◽

Cited By ~ 10

Author(s):

Saul J. Santivañez ◽

Mary L. Rodriguez ◽

Silvia Rodriguez ◽

Yashuito Sako ◽

Agathe Nkouawa ◽

...

Keyword(s):

Cystic Echinococcosis ◽

High Specificity ◽

Recombinant Antigen ◽

Enzyme Linked Immunosorbent Assay ◽

Parasitic Infections ◽

Clinical Settings ◽

Immunochromatographic Test ◽

Serum Samples ◽

Content Type ◽

Elisa Format

Diagnosis of cystic echinococcosis (CE) is based on the identification of the cyst(s) by imaging, using immunodiagnostic tests mainly as complementary tools in clinical settings. Among the antigens used for immunodiagnosis, previous studies described a good performance of the recombinant antigen B8/1 (rAgB) in an enzyme-linked immunosorbent assay (ELISA) format; however, in remote parts of areas where the disease is endemic, the implementation of an ELISA is difficult, so a more simple, rapid, and reliable method such as the immunochromatographic test (ICT) is required. In this study, using a set of 50 serum samples from patients with surgically confirmed CE, we compared the performance of an ICT and that of an ELISA using the rAgB. The overall sensitivities of ICT and ELISA were not statistically different (78% versus 72%;P= 0.36). The overall agreement between both tests was moderate (κ = 0.41;P< 0.01). Concordance between ICT and ELISA was substantial or almost perfect for patients with liver involvement (κ = 0.65;P< 0.001) and patients with more than one hydatid cyst (κ = 0.82;P< 0.001), respectively. Moreover, specificity analysis using a total of 88 serum samples from healthy individuals (n= 20) and patients (n= 68) with other parasitic infections revealed that ICT had a specificity of 89.8%. ICT and ELISA had similar performance for the detection of specific antibodies toE. granulosus, and ICT had a high specificity, opening the possibility of using ICT as a screening tool in rural settings.

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Detection of Francisella tularensis-Specific Antibodies in Patients with Tularemia by a Novel Competitive Enzyme-Linked Immunosorbent Assay

Clinical and Vaccine Immunology ◽

10.1128/cvi.00516-12 ◽

2012 ◽

Vol 20 (1) ◽

pp. 9-16 ◽

Cited By ~ 28

Author(s):

Neekun Sharma ◽

Akitoyo Hotta ◽

Yoshie Yamamoto ◽

Osamu Fujita ◽

Akihiko Uda ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Francisella Tularensis ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

The Novel ◽

Serum Samples ◽

Content Type ◽

Microagglutination Test ◽

Highly Sensitive ◽

High Degree

ABSTRACTA novel competitive enzyme-linked immunosorbent assay (cELISA) was developed and evaluated for detection of antibodies againstFrancisella tularensisin humans. The assay is based on the ability of serum antibodies to inhibit the binding of monoclonal antibodies (MAbs) directed againstF. tularensislipopolysaccharide antigens. The assay was evaluated using serum samples of tularemia patients, inactivatedF. tularensis-immunized rabbits, andF. tularensis-infected mice. Antibodies againstF. tularensiswere successfully detected in serum samples of tularemia patients as well as the immunized and infected animals. The cELISA method was compared to indirect ELISA (iELISA) and the commonly used microagglutination test (MA) using serum samples of 19 tularemia patients and 50 healthy individuals. The sensitivity and specificity of cELISA were 93.9 and 96.1%, respectively, in comparison to the iELISA. MA was less sensitive than cELISA with a sensitivity and specificity of only 81.8 and 98.0%, respectively. A high degree of correlation (R2= 0.8226) was observed between cELISA and iELISA results. The novel cELISA developed in this study appears to be highly sensitive and specific for serodiagnosis of human tularemia. The potential of the MAb-based cELISA to be used in both human and animal samples emphasizes its usefulness for serological survey of tularemia among multiple animal species.

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Evaluation of the performance of three serological tests for diagnosis of Leishmania infantum infection in dogs using latent class analysis

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612020105 ◽

2020 ◽

Vol 29 (4) ◽

Author(s):

Asier Basurco ◽

Alda Natale ◽

Katia Capello ◽

Antonio Fernández ◽

María Teresa Verde ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Leishmania Infantum ◽

Latent Class ◽

Rapid Test ◽

Antibody Test ◽

Latent Class Models ◽

Canine Leishmaniasis ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Serological Tests

Abstract Canine leishmaniasis (CanL) is a disease caused by Leishmania infantum. Serological methods are the most common diagnostic techniques used for the diagnosis of the CanL. The objective of our study was to estimate the sensitivity and specificity of one in-house ELISA kit (ELISA UNIZAR) and three commercially available serological tests (MEGACOR Diagnostik GmbH) including an immunochromatographic rapid test (FASTest LEISH®), an immunofluorescent antibody test (MegaFLUO LEISH®) and an enzyme-linked immunosorbent assay (MegaELISA LEISH®), using latent class models in a Bayesian analysis. Two hundred fifteen serum samples were included. The highest sensitivity was achieved for FASTest LEISH® (99.38%), ELISA UNIZAR (99.37%), MegaFLUO LEISH® (99.36%) followed by MegaELISA LEISH® (98.49%). The best specificity was obtained by FASTest LEISH® (98.43%), followed by ELISA UNIZAR (97.50%), whilst MegaFLUO LEISH® and MegaELISA LEISH® obtained the lower specificity (91.94% and 91.93%, respectively). The results of present study indicate that the immunochromatographic rapid test evaluated FASTest LEISH® show similar levels of sensitivity and specificity to the quantitative commercial tests. Among quantitative serological tests, sensitivity and specificity were similar considering ELISA or IFAT techniques.

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