Seroprevalence of Antibodies to Bartonella henselae in Patients with Cat Scratch Disease and in Healthy Controls: Evaluation and Comparison of Two Commercial Serological Tests

ABSTRACT Serologic testing for the presence of antibodies toBartonella henselae is a widely accepted diagnostic procedure for laboratory confirmation of the diagnosis of cat scratch disease (CSD). In this study a commercially available indirect immunofluorescence assay (IFA) based on B. henselae-infected human larynx carcinoma cells (test A) was evaluated. Sera from 42 patients with CSD (20 confirmed by PCR) and 270 sera from healthy controls (consisting of 63 cat owners, 65 individuals whose last close contact with cats was >6 months previously, and 142 persons who had never been exposed to cats) were investigated for antibodies to B. henselae. All patients with CSD had titers of immunoglobulin G (IgG) to B. henselae of 128 or higher (test A; sensitivity, 100%). Of the 270 controls 189 (70%) were seronegative (titer, <64), 38 (14.1%) had titers of 64, 30 (11.1%) had titers of 128, 9 (3.3%) had titers of 256, and 4 (1.5%) had high titers, 512 (test A; specificity, 70%). Of the cat owners and individuals who had never had close contact with cats, 71.4 and 71.12%, respectively, were seronegative, and titers of 64, 128, 256, and 512 were found in 14.3 and 16.2%, 1.6 and 10.5%, 9.5 and 0.7%, and 3.2 and 1.4%, respectively. The sera from the patients and from the first 100 healthy adults without a history of close contact with cats were additionally tested with a second commercially available IFA, based on Vero cells infected withB. henselae and Bartonella quintana (test B). The sensitivity and specificity of test B were 93 and 73%, respectively. For patients with CSD the cross-reactivity betweenB. henselae and B. quintana in this test was 95%. Both systems are highly sensitive but less specific for detection of IgG antibodies to B. henselae in samples from patients with clinically apparent CSD. For detection of IgM antibodies, test A seems to be more sensitive (88%) and more specific (95%) than test B (sensitivity and specificity of 64 and 86%, respectively). The data show that the seroprevalence of antibodies toB. henselae in German individuals is high (30%). Low antibody levels are not sufficient evidence of active or prior infection.

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Western Immunoblotting for Bartonella Endocarditis

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.10.1.95-102.2003 ◽

2003 ◽

Vol 10 (1) ◽

pp. 95-102 ◽

Cited By ~ 37

Author(s):

Pierre Houpikian ◽

Didier Raoult

Keyword(s):

Western Blot ◽

Cross Reactivity ◽

Molecular Techniques ◽

Cat Scratch Disease ◽

Serum Samples ◽

Western Immunoblotting ◽

Serological Tests ◽

Heterologous Antigen ◽

Bartonella Quintana ◽

Causative Species

ABSTRACT To differentiate infectious endocarditis (IE) from other Bartonella infections and to identify infecting Bartonella bacteria at the species level on a serological basis, we used Western immunoblotting to test sera from 51 patients with Bartonella IE (of which 27 had previously benefited from species identification by molecular techniques), 11 patients with chronic Bartonella quintana bacteremia, and 10 patients with cat scratch disease. Patients with IE were Western blot positive in 49 of 51 cases, and significant cross-reactivity with three heterologous Bartonella antigens was found in 45 of 49 cases. Sera from bacteremic patients did not react with more than one heterologous antigen, and sera from patients with cat scratch disease gave negative results. Sera reacted only with B. henselae in four cases of IE, including one with a positive PCR result for valve tissue. Western blot and cross-adsorption performed on serum samples from patients with IE (the identity of the causative species having been determined by PCR) were demonstrated to identify efficiently the causative species in all cases. When applied to patients diagnosed on the basis of serological tests only, this technique allowed identification of the causative species in 20 of 22 cases. The results were in accordance with epidemiological features. Six reactive bands of B. quintana (of molecular sizes from 10 to 83 kDa) demonstrated significant association with sera from patients with B. quintana endocarditis. Overall, Western blotting and cross-adsorption made it possible to identify the causative species in 49 of 51 (96%) IE cases.

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Characterization of Human Immunoglobulin (Ig) Isotype and IgG Subclass Response to Bartonella henselae Infection

Infection and Immunity ◽

10.1128/iai.66.12.5915-5920.1998 ◽

1998 ◽

Vol 66 (12) ◽

pp. 5915-5920 ◽

Cited By ~ 28

Author(s):

Svena L. McGill ◽

Russell L. Regnery ◽

Kevin L. Karem

Keyword(s):

Western Blot ◽

Blot Analysis ◽

Western Blot Analysis ◽

Bartonella Henselae ◽

Negative Control ◽

Cross Reactivity ◽

Antibody Activity ◽

Igg Subclass ◽

Banding Patterns ◽

Bartonella Quintana

ABSTRACT Serologic parameters of cat scratch disease (CSD) were evaluated by Western blot analysis. Sera from patients with serologically confirmed CSD antigen were screened for immunoglobulin (Ig) isotype-specific as well as IgG subclass-specific reactivity against Bartonella henselae whole-cell antigen. Bartonella-negative control sera were used to determine baseline antibody activity. Heterogeneous B. henselae-specific IgG reactivity with numerous protein bands, ranging from >150 to <17 kDa, was observed. Though individual banding patterns were variable, one approximately 83-kDa B. henselae protein (Bh83) was immunoreactive with all CSD sera tested, suggesting it is a conserved antigen during infection. Bh83 was not recognized by reference human antisera againstRickettsia rickettsii, Chlamydia group positive, Treponema pallidum, Orientia tsutsugamushi, Fransciscella tularensis,Ehrlichia chaffeensis, Mycoplasma pneumoniae, and Escherichia coli, although other cross-reactive proteins were evident. Significantly, CSD sera failed to recognize the 83-kDa protein when tested against Bartonella quintanaantigen, though sera from B. quintana-infected patients did react to Bh83. This cross-reactivity suggests epitope conservation during infection with B. henselae or B. quintana. Western blot analysis further revealed similar banding patterns when B. henselae was reacted against the Ig isotypes IgG and IgG1 and both secretory and alpha chains of IgA. Neither IgM nor IgE reacted significantly toBartonella antigen by our Western blot analysis. Dissection of the antibody response at the IgG subclass level indicated that prominent antigen recognition was limited to IgG1. These observations provide insight into induced immunity during CSD and provide evidence for conserved epitope expression during infection withB. henselae or B. quintana.

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The Bartonella henselae sucB gene encodes a dihydrolipoamide succinyltransferase protein reactive with sera from patients with cat-scratch disease

Journal of Medical Microbiology ◽

10.1099/jmm.0.45616-0 ◽

2004 ◽

Vol 53 (12) ◽

pp. 1221-1227 ◽

Cited By ~ 8

Author(s):

Christine M Litwin ◽

Joel M Johnson ◽

Thomas B Martins

Keyword(s):

Coxiella Burnetii ◽

Brucella Melitensis ◽

Bartonella Henselae ◽

Immunoblot Analysis ◽

Cross Reactivity ◽

Cosmid Library ◽

Cat Scratch Disease ◽

Igg Antibodies ◽

Bacillary Angiomatosis ◽

Dihydrolipoamide Succinyltransferase

Bartonella henselae is a recently recognized pathogenic bacterium associated with cat-scratch disease, bacillary angiomatosis and bacillary peliosis. A recombinant clone expressing an immunoreactive antigen of B. henselae was isolated by screening a genomic DNA cosmid library by Western blotting with sera pooled from patients positive for B. henselae IgG antibodies by indirect immunofluorescence (IFA). The deduced amino acid sequence of the 43.7 kDa encoded protein was found to be 76.3 % identical to the dihydrolipoamide succinyltransferase enzyme (SucB) of Brucella melitensis. SucB has been shown to be an immunogenic protein during infections by Brucella melitensis, Coxiella burnetii and Bartonella vinsonii. The agreement between reactivity with a recombinant SucB fusion protein on immunoblot analysis and the results obtained by IFA was 55 % for IFA-positive sera and 88 % for IFA-negative sera. Cross-reactivity was observed with sera from patients with antibodies against Brucella melitensis, Mycoplasma pneumoniae, Francisella tularensis, Coxiella burnetii and Rickettsia typhi.

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Cat Scratch Disease: The Story Continues

Canadian Journal of Infectious Diseases ◽

10.1155/1997/982908 ◽

1997 ◽

Vol 8 (1) ◽

pp. 43-49 ◽

Cited By ~ 6

Author(s):

Mary Anne Opavsky

Keyword(s):

Clinical Characteristics ◽

Bartonella Henselae ◽

Data Extraction ◽

Significant Risk ◽

Unknown Origin ◽

Etiological Agent ◽

Cat Scratch Disease ◽

Serological Tests ◽

Medline Search ◽

Atypical Presentations

OBJECTIVE: To present a perspective on the current state of knowledge of cat scratch disease (CSD), including the evidence forBartonella henselaeas the etiological agent, epidemiological and clinical characteristics of the disease, available diagnostic tests and current therapeutic options.DATA SOURCES: MEDLINE search of the literature published from 1966 to 1995 using ‘cat scratch disease’, ‘Bartonella henselae’, ‘Rochalimaea henselae’ as key words and bibliographies of selected papers.DATA EXTRACTION: Selected studies reporting data on etiology, epidemiology, clinical characteristics, diagnosis and therapy of CSD were evaluated.DATA SYNTHESIS AND CONCLUSIONS: Evidence accumulated to date supportsB henselaeas the etiological agent of CSD. The most significant risk factors for CSD are being licked on the face, scratched or bitten by a kitten and owning a kitten with fleas. Available serological tests can confirm classic CSD and identifyB henselaeas the cause of more atypical presentations, such as fever of unknown origin, granulomatous hepatitis, encephalitis and osteomyelitis. Symptomatic management is appropriate for isolated lymphadenopathy caused by CSD in healthy individuals; however, antibiotic therapy may be indicated for patients with more severe manifestations of the disease and immunocompromised hosts. Further study of CSD, in particular the epidemiology and therapy, is warranted. A better understanding of the pathogenesis ofB henselaeinfection will have important implications in both immunocompetent and immunocompromised individuals.

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THU0310 CASE–CONTROL SEROPREVALENCE STUDY ON THE ASSOCIATION BETWEEN BARTONELLA INFECTION AND ANTI-NEUTROPHIL CYTOPLASMIC ANTIBODY-ASSOCIATED VASCULITIS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.3361 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 384.1-384

Author(s):

A. Mahr ◽

S. Edouard ◽

D. Cornec ◽

S. Gonzalez-Chiappe ◽

J. Goronzy ◽

...

Keyword(s):

Bartonella Henselae ◽

The United States ◽

Cross Reactivity ◽

Endothelial Damage ◽

Nasal Discharge ◽

Healthy Controls ◽

Serological Testing ◽

Research Support ◽

Positive Serology ◽

Organ Systems

Background:Bartonellosis is an emerging anthropozoonosis caused by infection with intracellular Gram-negativeBartonellaspecies. It leads to necrotizing granulomas and endothelial damage and causes acute and chronic human diseases, such as cat scratch disease, bacillary angiomatosis and endocarditis. Endocarditis due toBartonella henselaeandB. quintanais reported to produce anti-neutrophil cytoplasmic antibodies (ANCAs) that disappear with effective antimicrobial treatment.Objectives:Hypothesizing a role forBartonellainfection in ANCA-associated vasculitis (AAV), which also includes granulomatous and vascular inflammation, we studied the seroprevalence of 5Bartonellaspecies in patients with AAV.Methods:The study used plasma samples from patients with granulomatosis with polyangiitis and microscopic polyangiitis that were enrolled in the Rituximab for AAV (RAVE) trial and from healthy controls living in the United States. Western blot assays were used for serological testing of infection withB. quintana,B. henselaeHouston-1,B. elizabethae,B. vinsoniisubsp.berkhoffiiandB. alsatica. The associations of positive serology results and AAV were expressed as odds ratios (OR). Clinical characteristics of seropositive and seronegative patients, assessed by the BVAS/WG instrument, were compared. These comparisons were done for 9 organ systems; in case they showed differences withP<0.10, the corresponding organ system-specific clinical features were also analyzed. Statistical analysis was performed using Fisher’s exact test or Student’s t-test, as appropriate.Results:We analyzed blood samples of 187 patients with AAV (collected at start of the trial) and of 127 controls. There were no significant differences between the cases and controls for mean age (P=0.148) and proportion of males (P=0.36).Bartonellaspp. serological testing was positive for 112 (60%) cases and 40 (31%) controls (OR 3.25 [95% CI 2.02–5.22],P<0.001). Significant associations were also found within subsets of PR3-AAV (OR 4.00 [95% CI 2.37–6.76],P<0.001), MPO-AAV (OR 2.18 [95% CI 1.17–4.06],P=0.017), newly-diagnosed (OR 3.89 [95% CI 2.21–6.86],P<0.001) and relapsing disease (OR 2.86 [95% CI 1.65–4.98],P<0.001). Species-specific positive serological testing was found in particular againstB. henselae(cases: 27%, controls: 0.8%; OR 39.93 [95% CI 5.42–293.90];P<0.001). Compared to AAV patients without seropositivity forBartonellaspp., AAV patients testing seropositive forBartonellaspp. had significantly more bloody nasal discharge (P=0.046), sinus involvement (P=0.035) and conjunctivitis/episcleritis (P=0.016).Conclusion:This study reveals higher seroprevalence ofBartonella, especiallyB. henselae, in patients with AAV than in healthy controls. Although cross-reactivity ofBartonellawith other microorganisms cannot be excluded, these results may support an etiopathogenic role ofBartonellainfection in AAV that deserves further investigation.Disclosure of Interests:Alfred Mahr Consultant of: Celgene, Speakers bureau: Roche, Chugai, Sophie Edouard: None declared, Divi Cornec: None declared, Solange GONZALEZ-CHIAPPE: None declared, Jörg Goronzy: None declared, Philippe Guilpain: None declared, Carol Langford: None declared, Pierre-Yves Lévy: None declared, Peter A. Merkel: None declared, Paul Monach: None declared, E. William St. Clair: None declared, Philip Seo: None declared, Robert Spiera Grant/research support from: Roche-Genetech, GSK, Boehringer Ingelheim, Chemocentryx, Corbus, Forbius, Sanofi, Inflarx, Consultant of: Roche-Genetech, GSK, CSL Behring, Sanofi, Janssen, Chemocentryx, Forbius, Mistubishi Tanabe, Cornelia Weyand: None declared, John H. Stone Grant/research support from: Roche, Consultant of: Roche, Didier Rauolt: None declared, Ulrich Specks: None declared

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Cat scratch disease as zoonosis: Pathogenesis, clinical symptoms, diagnosis.

Medycyna Weterynaryjna ◽

10.21521/mw.5997 ◽

2018 ◽

Vol 74 (1) ◽

pp. 5997-2018

Author(s):

ŁUKASZ MAZUREK ◽

STANISŁAW WINIARCZYK ◽

ŁUKASZ ADASZEK

Keyword(s):

Clinical Symptoms ◽

Bartonella Henselae ◽

Unknown Origin ◽

Skin Lesions ◽

Cat Scratch Disease ◽

Serological Tests ◽

Antibody Titres ◽

Polymerase Chain ◽

Brain And Spinal Cord ◽

The Brain

The cat scratch disease in humans is caused by the bacteria Bartonella henselae. The disease can take many different forms: from asymptomatic cases, cases of skin lesions, fever of unknown origin, enlargement of lymph nodes, ophthalmologic disorders, to severe cases involving inflammation of the brain and spinal cord or endocarditis. The reservoir of B. henselae for humans are domestic animals, especially cats. The diagnosis of the disease is based on data from the anamnesis, the patient’s confirmed exposure to cats, and the results of serological tests showing an increase in antibody titres for B. henselae. The disease can also be confirmed by positive results of the polymerase chain reaction (PCR). No vaccines against bartonellosis are available. The most important in preventing the disease is to maintain appropriate hygiene in contact with cats and dogs, and to eradicate the vectors of Bartonella, such as fleas..

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#28: Rapid, Non-invasive Detection of Invasive Bartonella Infections in Pediatric Patients Using the Karius Test, A Next-Generation Sequencing Test for Microbial Cell-free DNA in Plasma

Journal of the Pediatric Infectious Diseases Society ◽

10.1093/jpids/piab031.023 ◽

2021 ◽

Vol 10 (Supplement_2) ◽

pp. S11-S11

Author(s):

Nicholas Degner ◽

Matt Smollin ◽

Ozlem Equils ◽

Aparna Arun ◽

Christiaan DeVries ◽

...

Keyword(s):

Prosthetic Valve ◽

Fever Of Unknown Origin ◽

Bartonella Henselae ◽

Prosthetic Valve Endocarditis ◽

Unknown Origin ◽

Cat Scratch Disease ◽

Bartonella Quintana ◽

Non Invasive ◽

Free Dna ◽

Generation Sequencing

Abstract Background Bartonella henselae and Bartonella quintana are the etiologic agents of cat scratch disease (CSD) and “trench fever”, respectively. Both are important causes of culture-negative endocarditis and fever of unknown origin (FUO). The diagnosis of Bartonella infections is limited by (1) the nonspecific, protean manifestations of the disease and its broad differential diagnosis; (2) the fastidious nature of Bartonella spp., leading to rare detections with traditional culture based methods; (3) the insensitivity and poor specificity of Bartonella serologies. Rapid, non-invasive diagnosis of Bartonella through next-generation sequencing (NGS) of plasma microbial cell-free DNA (mcfDNA) offers a means to overcome these limitations. Here we describe the diagnosis of 23 Bartonella infections in children from August 2017 – December 2020 using plasma mcfDNA NGS. Methods The Karius Test (KT), developed and validated in Karius’ CLIA certified/CAP accredited lab, detects mcfDNA in plasma. After mcfDNA is extracted and NGS performed, human reads are removed, and remaining sequences are aligned to a curated database of > 1400 organisms. McfDNA from organisms present above a statistical threshold are reported and quantified in molecules/μL (MPM). Clinical information was included from data submitted with the requisition or obtained at the time of reporting from clinical consultations with the provider. Results KT detected Bartonella henselae mcfDNA in 22 cases and Bartonella quintana in 1. Detections included 10 cases of endocarditis (7 prosthetic valve), 12 cases of CSD/FUO, and a single case of osteomyelitis. Glomerulonephritis was reported in 5 the cases of endocarditis. Six cases had splenic involvement; three had hepatic involvement. History of cat exposure was elicited in 8 cases. The mean MPMs was highest for prosthetic valve endocarditis (mean 47,272 +/- 67,526) followed by native valve endocarditis (3,881 +/- 2,458), FUO/CSD (1,922 +/- 3,416), and osteomyelitis (119 +/- 0) (p<0.05). Three subjects had serial mcfDNA monitoring. Predictable declines in Bartonella mcfDNA were observed in response to therapy in all three patients. The duration of positive Bartonella mcfDNA signal ranged from 22–42 days (30.7, +/- 10.3). Conclusion Open-ended, plasma-based NGS for mcfDNA provides a rapid, non-invasive method to diagnose diverse clinical manifestations of invasive pediatric Bartonella infection against a competing broad differential diagnosis. The quantification of mcfDNA may further help in differentiating the various clinical syndromes caused by Bartonella. Finally, serial monitoring to trend MPMs may serve as an indicator of burden of infection, provide a means to monitor treatment efficacy and ultimately help define the length of therapy for optimal outcomes. All detections are Bartonella henselae except for one case of *Bartonella quintana (prosthetic valve endocarditis). MPM=molecules/μL; GN=glomerulonephritis, LN=lymph node involvement, FUO=fever of unknown origin, CSD=cat scratch disease

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Comparison of In-House and Commercial Slides for Detection by Immunofluorescence of Immunoglobulins G and M against Bartonella henselae and Bartonella quintana

Clinical and Vaccine Immunology ◽

10.1128/cdli.9.5.1004-1009.2002 ◽

2002 ◽

Vol 9 (5) ◽

pp. 1004-1009 ◽

Cited By ~ 7

Author(s):

M. Maurin ◽

J. M. Rolain ◽

D. Raoult

Keyword(s):

Bartonella Henselae ◽

Fluorescent Antibody ◽

The Other ◽

Cat Scratch Disease ◽

Indirect Fluorescent Antibody ◽

Igg Antibodies ◽

Serum Samples ◽

Bartonella Quintana ◽

Antibody Tests ◽

Higher Sensitivity

ABSTRACT We compared the sensitivities and specificities of indirect fluorescent antibody tests developed in our laboratory and commercially available from Focus Technologies (FT; formerly MRL Diagnostic) for detection of serum antibodies to Bartonella spp. Serum samples tested were from patients with culture- or PCR-confirmed Bartonella quintana or B. henselae infections causing cat scratch disease (CSD), chronic bacteremia, or endocarditis. At a cutoff titer of 64, the FT test had higher sensitivity than our in-house test in detecting anti-B. henselae immunoglobulin G (IgG) antibodies in CSD patients (91.2 versus 52.9%; P < 0.001). The specificity in serum samples from 85 control patients was, however, lower with the FT test (87%) than with the in-house test (98.8%) (P = 0.002). A cutoff titer of 128 improves the specificity for the FT test but lowers the sensitivity to 85%. For patients infected with B. henselae, our in-house test, but not the FT test, enabled endocarditis to be detected more reliably. With the in-house test, titers of IgG against B. henselae of ≥1,024 were found only in endocarditis patients and not in CSD patients. With the FT test, 19.1% of CSD patients had titers of IgG against B. henselae of ≥1,024 (P < 0.001). Our in-house technique also improved detection of anti-B. quintana antibodies in homeless patients with endocarditis. IgG titers of ≥1,024 were present in 75% of serum samples, but only in 16.7% of serum samples with the FT test (P = 0.004). Since each test has advantages over the other, the serological diagnosis of Bartonella infections would benefit if both tests were used concurrently.

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A new ELISA and western blot technique based on recombinant TES antigen and/or larval antigen for the detection of toxocariasis in humans

Parasitology ◽

10.1017/s0031182020002085 ◽

2020 ◽

pp. 1-8

Author(s):

Marie-Kristin Raulf ◽

Daniela Jordan ◽

Herbert Auer ◽

Jens M. Warnecke ◽

Bernd Lepenies ◽

...

Keyword(s):

Sensitivity And Specificity ◽

High Reliability ◽

High Sensitivity ◽

Cross Reactivity ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Serological Tests ◽

Western Blots ◽

Blot Technique ◽

Human Sera

Abstract Serological antibody detection by enzyme-linked immunosorbent assay (ELISA)- and immunoblot-based methods constitutes the best indicator of human Toxocara infection. Nevertheless, the availability of serological tests, particularly western blots (WB), evaluated for sensitivity and specificity is limited. Therefore, an Anti-Toxocara-ELISA immunoglobulin g (IgG) prototype (Proto-ELISA) and an Anti-Toxocara-Westernblot (IgG) prototype (Proto-WB) were evaluated by testing 541 human sera pre-determined for Toxocara infection by an established in-house Anti-Toxocara-ELISA (IH-ELISA). To evaluate sensitivity and specificity of the newly developed ELISA and WB prototypes, results were compared to IH-ELISA and a commercial WB (Com-WB). Compared to the IH-ELISA, a sensitivity of 93.1% (229/246) and a specificity of 94.6% (279/295) of the Proto-ELISA with a Cohen's κ of 0.88 were obtained. The sensitivity of the Proto-WB was 76.7% (240/313) and specificity was 99.6% (227/228) with a Cohen's κ of 0.73 compared to those of Com-WB. A comparison to the IH-ELISA revealed 91.5% (225/246) sensitivity and 94.6% (279/295) specificity of the Proto-WB with a Cohen's κ of 0.86. Cross-reactivity was observed for some samples positive for Ascaris and Trichinella spp. in the Proto-ELISA, Proto-WB and Com-WB. Overall, the evaluated ELISA and WB prototypes showed high sensitivity and specificity, indicating high reliability of these newly developed tests.

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