Field Application of Serodiagnostics To Identify Elephants with Tuberculosis prior to Case Confirmation by Culture

ABSTRACTThree serologic methods for antibody detection in elephant tuberculosis (TB), the multiantigen print immunoassay (MAPIA), ElephantTB STAT-PAK kit, and DPP VetTB test, were evaluated using serial serum samples from 14 captive elephants infected withMycobacterium tuberculosisin 5 countries. In all cases, serological testing was performed prior to the diagnosis of TB by mycobacterial culture of trunk wash or tissue samples collected at necropsy. All elephants produced antibody responses toM. tuberculosisantigens, with 13/14 recognizing ESAT-6 and/or CFP10 proteins. The findings supported the high serodiagnostic test accuracy in detecting infections months to years beforeM. tuberculosiscould be isolated from elephants. The MAPIA and/or DPP VetTB assay demonstrated the potential for monitoring antimycobacterial therapy and predicting TB relapse in treated elephants when continuously used in the posttreatment period. History of exposure to TB and past treatment information should be taken into consideration for proper interpretation of the antibody test results. Data suggest that the more frequent trunk wash culture testing of seropositive elephants may enhance the efficiency of the TB diagnostic algorithm, leading to earlier treatment with improved outcomes.

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Help in the Choice of Automated or Semiautomated Immunoassays for Serological Diagnosis of Toxoplasmosis: Evaluation of Nine Immunoassays by the French National Reference Center for Toxoplasmosis

Journal of Clinical Microbiology ◽

10.1128/jcm.01193-16 ◽

2016 ◽

Vol 54 (12) ◽

pp. 3034-3042 ◽

Cited By ~ 11

Author(s):

O. Villard ◽

B. Cimon ◽

C. L'Ollivier ◽

H. Fricker-Hidalgo ◽

N. Godineau ◽

...

Keyword(s):

Congenital Toxoplasmosis ◽

Antibody Detection ◽

Clinical Settings ◽

Serum Samples ◽

Igg Antibody ◽

Routine Testing ◽

Content Type ◽

Specificity And Sensitivity ◽

National Reference Center ◽

Serology Testing

Toxoplasmosis, a benign infection, is asymptomatic or paucisymptomatic in over 80% of cases, except in immunocompetent patients suffering from ocular toxoplasmosis or in immunocompromised patients with opportunistic or congenital toxoplasmosis. Diagnosis is based mainly on serology testing. Thus, we compared the performance of the nine most commonly used commercial automated or semiautomated immunoassays for IgG and IgMToxoplasma gondiiantibody detection, that is, the Advia Centaur, Architect, AxSYM, Elecsys, Enzygnost, Liaison, Platelia, VIDAS, and VIDIA assays. The assays were conducted on four panels of serum samples derived during routine testing from patients with an interfering disease and who exhibited a low IgG antibody level in one of two clinical settings, namely, acute or chronic toxoplasmosis. As a result, IgG sensitivities ranged from 97.1% to 100%, and IgG specificities ranged from 99.5% to 100%. For IgG quantification, strong differences in IgG titers (expressed in IU/ml) were noted depending on the assay used. IgM sensitivities ranged from 65% to 97.9%, and IgM specificities ranged from 92.6% to 100%. For defining the best serological strategies to be implemented, it appears crucial to compare the diagnostic performance of the different tests with respect to their specificity and sensitivity in detecting the presence of IgG and IgM antibodies.

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Prevalence and genetic characterization of Toxoplasma gondii in naturally infected backyard pigs intended for familial consumption in Romania

Parasites & Vectors ◽

10.1186/s13071-019-3842-8 ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 2

Author(s):

Anamaria Ioana Paştiu ◽

Anamaria Cozma-Petruț ◽

Aurélien Mercier ◽

Anamaria Balea ◽

Lokman Galal ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Rural Areas ◽

Antibody Test ◽

Antibody Detection ◽

Serum Samples ◽

Potential Threat ◽

Polymerase Chain ◽

Pcr Targeting ◽

Backyard Pigs

Abstract Background Foodborne toxoplasmosis in humans can be due to the exposure to tissue cysts of Toxoplasma gondii through the consumption of meat, including pork, of infected animals. Traditional Romanian food habits include pork as the preferred meat, while backyard pig rearing remains a common practice in many rural areas of Romania. The aims of the present study were to estimate the prevalence of T. gondii infection in naturally infected backyard pigs slaughtered for familial consumption and to genetically characterize the T. gondii strains obtained. Methods Paired blood and heart samples were collected from 94 backyard pigs, home slaughtered for private consumption. Serum samples were analyzed using the immunofluorescence antibody test (IFAT) for anti-T. gondii antibody detection. Heart samples were screened by polymerase chain reaction (PCR) targeting the 529-bp repeat region (REP529) for T. gondii detection. In addition, heart samples from IFAT positive animals were bioassayed in mice. The T. gondii isolates were genotyped by the analysis of 15 microsatellite markers. Results The results showed that almost half of the pigs investigated were T. gondii seropositive (46.8%, 95% confidence interval (CI): 36.4–57.4%) and in more than a quarter of the pigs (26.6%, 95% CI: 18.0–36.7%), the parasite was detected by PCR. Three (3/44) T. gondii strains were isolated from hearts of seropositive pigs and they all belonged to genotype II. Conclusions The present study showed the presence of T. gondii infection in backyard pigs in Romania, which suggests that consumption of pork from animals reared and slaughtered at home may pose a potential threat to human health and should be given attention. In addition, to our knowledge, this is the first study to provide data concerning T. gondii strains circulating in pigs from Romania.

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Rapid Detection of Serum Antibody by Dual-Path Platform VetTB Assay in White-Tailed Deer Infected with Mycobacterium bovis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00120-13 ◽

2013 ◽

Vol 20 (6) ◽

pp. 907-911 ◽

Cited By ~ 18

Author(s):

Konstantin P. Lyashchenko ◽

Rena Greenwald ◽

Javan Esfandiari ◽

Daniel J. O'Brien ◽

Stephen M. Schmitt ◽

...

Keyword(s):

Diagnostic Accuracy ◽

Mycobacterium Bovis ◽

Skin Testing ◽

Serum Antibody ◽

Antibody Responses ◽

Antibody Detection ◽

Serum Samples ◽

Content Type ◽

In The Wild ◽

Free Ranging

ABSTRACTBovine tuberculosis (TB) in cervids remains a significant problem affecting farmed herds and wild populations. Traditional skin testing has serious limitations in certain species, whereas emerging serological assays showed promising diagnostic performance. The recently developed immunochromatographic dual-path platform (DPP) VetTB assay has two antigen bands, T1 (MPB83 protein) and T2 (CFP10/ESAT-6 fusion protein), for antibody detection. We evaluated the diagnostic accuracy of this test by using serum samples collected from groups of white-tailed deer experimentally inoculated withMycobacterium bovis,M. aviumsubsp.paratuberculosis, orM. bovisBCG Pasteur. In addition, we used serum samples from farmed white-tailed deer in herds with no history of TB, as well as from free-ranging white-tailed deer culled during field surveillance studies performed in Michigan known to have bovine TB in the wild deer population. The DPP VetTB assay detected antibody responses in 58.1% of experimentally infected animals within 8 to 16 weeks postinoculation and in 71.9% of naturally infected deer, resulting in an estimated test sensitivity of 65.1% and a specificity of 97.8%. The higher seroreactivity found in deer with naturally acquiredM. bovisinfection was associated with an increased frequency of antibody responses to the ESAT-6 and CFP10 proteins, resulting in a greater contribution of these antigens, in addition to MPB83, to the detection of seropositive animals, compared with experimentalM. bovisinfection. Deer experimentally inoculated with eitherM. aviumsubsp.paratuberculosisorM. bovisBCG Pasteur did not produce cross-reactive antibodies that could be detected by the DPP VetTB assay. The present findings demonstrate the relatively high diagnostic accuracy of the DPP VetTB test for white-tailed deer, especially in the detection of naturally infected animals.

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Lung Tissue Concentrations of Pyrazinamide among Patients with Drug-Resistant Pulmonary Tuberculosis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00226-17 ◽

2017 ◽

Vol 61 (6) ◽

Cited By ~ 26

Author(s):

Russell R. Kempker ◽

M. Tobias Heinrichs ◽

Ketino Nikolaishvili ◽

Irina Sabulua ◽

Nino Bablishvili ◽

...

Keyword(s):

Pulmonary Tuberculosis ◽

Lung Tissue ◽

Ex Vivo ◽

Inverse Correlation ◽

Drug Resistant ◽

Tissue Penetration ◽

Serum Samples ◽

Tissue Samples ◽

Content Type ◽

Antituberculosis Treatment

ABSTRACT Improved knowledge regarding the tissue penetration of antituberculosis drugs may help optimize drug management. Patients with drug-resistant pulmonary tuberculosis undergoing adjunctive surgery were enrolled. Serial serum samples were collected, and microdialysis was performed using ex vivo lung tissue to measure pyrazinamide concentrations. Among 10 patients, the median pyrazinamide dose was 24.7 mg/kg of body weight. Imaging revealed predominant lung lesions as cavitary (n = 6 patients), mass-like (n = 3 patients), or consolidative (n = 1 patient). On histopathology examination, all tissue samples had necrosis; eight had a pH of ≤5.5. Tissue samples from two patients were positive for Mycobacterium tuberculosis by culture (pH 5.5 and 7.2). All 10 patients had maximal serum pyrazinamide concentrations within the recommended range of 20 to 60 μg/ml. The median lung tissue free pyrazinamide concentration was 20.96 μg/ml. The median tissue-to-serum pyrazinamide concentration ratio was 0.77 (range, 0.54 to 0.93). There was a significant inverse correlation between tissue pyrazinamide concentrations and the amounts of necrosis (R = −0.66, P = 0.04) and acid-fast bacilli (R = −0.75, P = 0.01) identified by histopathology. We found good penetration of pyrazinamide into lung tissue among patients with pulmonary tuberculosis with a variety of radiological lesion types. Our tissue pH results revealed that most lesions had a pH conducive to pyrazinamide activity. The tissue penetration of pyrazinamide highlights its importance in both drug-susceptible and drug-resistant antituberculosis treatment regimens.

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Enhanced Antibody Detection and Diagnosis of Coccidioidomycosis with the MiraVista IgG and IgM Detection Enzyme Immunoassay

Journal of Clinical Microbiology ◽

10.1128/jcm.01880-16 ◽

2017 ◽

Vol 55 (3) ◽

pp. 893-901 ◽

Cited By ~ 14

Author(s):

Joshua Malo ◽

Eric Holbrook ◽

Tirdad Zangeneh ◽

Chris Strawter ◽

Eyal Oren ◽

...

Keyword(s):

Enzyme Immunoassay ◽

Complement Fixation ◽

Community Acquired Pneumonia ◽

Antibody Detection ◽

Serum Samples ◽

Content Type ◽

Appropriate Treatment ◽

Detection And Diagnosis ◽

Disseminated Disease ◽

Improved Accuracy

ABSTRACT Coccidioidomycosis is a common cause of community-acquired pneumonia in areas of the southwestern United States in which the disease is endemic. Clinical presentations range from self-limited disease to severe disseminated disease. Therefore, early and accurate diagnosis is essential to ensure appropriate treatment and monitoring. Currently available diagnostic tests have variable accuracy, particularly in certain patient populations, and new tests may offer improved accuracy for the diagnosis of coccidioidomycosis. Serum samples from 103 cases of coccidioidomycosis and 373 controls were tested for IgG and IgM antibodies using the MVista anti- Coccidioides antibody enzyme immunoassay. Serum specimens from 170 controls from areas in which the disease is endemic and 44 cases were tested by immunodiffusion at MiraVista Diagnostics. The sensitivity of the MVista antibody assay was 88.3%, and the specificity was 90%. The sensitivity was maintained in the presence of immunocompromising conditions or immunosuppressive therapies. The sensitivity of immunodiffusion was 60.2%, and the specificity was 98.8%. The sensitivity of complement fixation (62 cases) was 66.1%, but the specificity could not be determined. The MVista anti- Coccidioides antibody enzyme immunoassay offers improved sensitivity, compared with immunodiffusion and complement fixation, is not impaired in immunocompromised patients, and permits highly reproducible semiquantification.

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Evaluation of Eight Serological Tests for Diagnosis of Imported Schistosomiasis

Clinical and Vaccine Immunology ◽

10.1128/cvi.05680-11 ◽

2012 ◽

Vol 19 (6) ◽

pp. 948-953 ◽

Cited By ~ 59

Author(s):

Hans-Friedemann Kinkel ◽

Sabine Dittrich ◽

Britta Bäumer ◽

Thomas Weitzel

Keyword(s):

Antibody Test ◽

Parasitic Infections ◽

Serum Samples ◽

Serological Tests ◽

Content Type ◽

Commercial Kits ◽

Few Data ◽

Indirect Immunofluorescent ◽

Positive Results ◽

Immunofluorescent Antibody Test

ABSTRACTThe diagnosis of schistosomiasis in individuals from countries where the disease is not endemic is challenging, and few data are available on the accuracy of serological diagnosis in those patients. We evaluated the performance of eight serological assays, including four commercial kits, in the diagnosis of imported schistosomiasis in individuals from areas where the disease is not endemic, including six enzyme-linked immunosorbent assays using three different antigens, an indirect hemagglutination assay, and an indirect immunofluorescent-antibody test. To analyze the assays, we used a total of 141 serum samples, with 121 derived from patients with various parasitic infections (among which were 37 cases of schistosomiasis) and 20 taken from healthy volunteers. The sensitivity values for detection of schistosomiasis cases ranged from 41% to 78% and were higher forSchistosoma mansonithan forS. haematobiuminfections. Specificity values ranged from 76% to 100%; false-positive results were most frequent for samples from patients with cestode infections. By combining two or more tests, sensitivity improved markedly and specificity decreased only moderately. Serological tests are useful instruments for diagnosing imported schistosomiasis in countries where the disease is not endemic, but due to limitations in test sensitivities, we recommend the use of two or more assays in parallel.

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Comparison of Three Immunoassays for Detection of Antibodies to Strongyloides stercoralis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00041-14 ◽

2014 ◽

Vol 21 (5) ◽

pp. 732-736 ◽

Cited By ~ 27

Author(s):

Neil W. Anderson ◽

Diane M. Klein ◽

Sarina M. Dornink ◽

Deborah J. Jespersen ◽

Joseph Kubofcik ◽

...

Keyword(s):

Strongyloides Stercoralis ◽

Antibody Detection ◽

Venn Diagram ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Perfect Agreement ◽

Content Type ◽

Culture Techniques ◽

Intestinal Nematode ◽

Diagram Analysis

ABSTRACTDue to the limited sensitivities of stool-based microscopy and/or culture techniques forStrongyloides stercoralis, the detection of antibodies to this intestinal nematode is relied upon as a surrogate for determining exposure status or making a diagnosis ofS. stercoralisinfection. Here, we evaluated three immunoassays, including the recently released InBios Strongy Detect IgG enzyme-linked immunosorbent assay (ELISA) (InBios International, Inc., Seattle, WA), the SciMedxStrongyloidesserology microwell ELISA (SciMedx Corporation, Denville, NJ), and the luciferase immunoprecipitation system (LIPS) assay performed at the National Institutes of Health (NIH), for their detection of IgG antibodies toS. stercoralis. A total of 101 retrospective serum samples, previously submitted for routineS. stercoralisantibody detection using the SciMedx assay, were also evaluated by the InBios and LIPS assays. The qualitative results from each assay were compared using a Venn diagram analysis, to the consensus result among the three assays, and each ELISA was also evaluated using the LIPS assay as the reference standard. By Venn diagram analysis, 65% (66/101) of the samples demonstrated perfect agreement by all three assays. Also, the numbers of samples considered positive or negative by a single method were similar. Compared to the consensus result, the overall percent agreement of the InBios, SciMedx, and LIPS assays were comparable at 87.1%, 84.2%, and 89.1%, respectively. Finally, the two ELISAs performed analogously but demonstrated only moderate agreement (kappa coefficient for the two assays, 0.53) with the LIPS assay. Collectively, while the two commercially available ELISAs perform equivalently, neither should be used independently of clinical evaluation to diagnose strongyloidiasis.

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Sensitive and Specific Enzyme-Linked Immunosorbent Assay for Detecting Serum Antibodies against Mycobacterium avium subsp. paratuberculosis in Fallow Deer

Clinical and Vaccine Immunology ◽

10.1128/cvi.00159-14 ◽

2014 ◽

Vol 21 (8) ◽

pp. 1077-1085 ◽

Cited By ~ 8

Author(s):

José M. Prieto ◽

Ana Balseiro ◽

Rosa Casais ◽

Naiara Abendaño ◽

Liam E. Fitzgerald ◽

...

Keyword(s):

Immunosorbent Assay ◽

Mycobacterium Avium ◽

Cost Effective ◽

Fallow Deer ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Igg Antibody ◽

Tissue Samples ◽

Content Type ◽

Mycobacterium Avium Subsp Paratuberculosis

ABSTRACTThe enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies againstMycobacterium aviumsubsp.paratuberculosisin wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deerM. aviumsubsp.paratuberculosisisolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detectM. aviumsubsp.paratuberculosisantibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations.

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Serological, molecular, and microscopic detection of Leishmania in cats (Felis catus) in Belo Horizonte, Minas Gerais State, Brazil

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-296120180052 ◽

2018 ◽

Vol 27 (4) ◽

pp. 570-574 ◽

Cited By ~ 8

Author(s):

Fernanda Morcatti Coura ◽

Stephanie Karoline Pereira Passos ◽

Marina de Oliveira França Pelegrino ◽

Fabiola de Oliveira Paes Leme ◽

Gustavo Fontes Paz ◽

...

Keyword(s):

Bone Marrow ◽

Antibody Test ◽

Canine Leishmaniasis ◽

Leishmania Infection ◽

Pcr Analysis ◽

Serum Samples ◽

Endemic Region ◽

Tissue Samples ◽

Human Leishmaniasis ◽

Belo Horizonte

Abstract The role of cats in the epidemiological cycle of leishmaniasis remains unclear. To better understand the occurrence of leishmaniasis in cats, we studied the frequency of Leishmania in serum samples of 100 cats living in an endemic region for canine and human leishmaniasis by serological, parasitological, and molecular methods. Of the 100 cats, 54 were seropositive for Leishmania antibodies by immunofluorescence antibody test. None of the bone marrow aspirates collected from these cats tested positive for the parasite in culture or upon polymerase chain reaction (PCR) analysis. Biopsy samples of the ears also tested negative for Leishmania upon PCR analysis. These findings may indicate that the region is endemic for canine leishmaniasis and cats are infected by Leishmania; or that cross-reaction with antibodies against other parasites increases the frequency of seropositivity; or that cats respond to Leishmania infection by producing antibodies when few or no parasites are present in bone marrow and tissue samples. Overall, our results suggest that cats can be infected by Leishmania ; however, we failed to demonstrate feline parasitosis. These findings highlight the need to study leishmaniasis in cats, since sandflies feed on cats, these animals may act as a reservoir for the parasite.

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Performance of Commercial Mycoplasma hyopneumoniae Serum Enzyme-Linked Immunosorbent Assays under Experimental and Field Conditions

Journal of Clinical Microbiology ◽

10.1128/jcm.00485-20 ◽

2020 ◽

Vol 58 (12) ◽

Author(s):

Ana Paula S. Poeta Silva ◽

Ronaldo L. Magtoto ◽

Henrique M. Souza Almeida ◽

Aric McDaniel ◽

Precy D. Magtoto ◽

...

Keyword(s):

Serum Antibody ◽

Mycoplasma Hyopneumoniae ◽

Error Rates ◽

False Positives ◽

Field Conditions ◽

Misclassification Error ◽

Antibody Detection ◽

Serum Samples ◽

Content Type ◽

Immunosorbent Assays

ABSTRACT Mycoplasma hyopneumoniae is an economically significant pathogen of swine. M. hyopneumoniae serum antibody detection via commercial enzyme-linked immunosorbent assays (ELISAs) is widely used for routine surveillance in commercial swine production systems. Samples from two studies were used to evaluate assay performance. In study 1, 6 commercial M. hyopneumoniae ELISAs were compared using serum samples from 8-week-old cesarean-derived, colostrum-deprived (CDCD) pigs allocated to the following 5 inoculation groups of 10 pigs each: (i) negative control, (ii) Mycoplasma flocculare (strain 27399), (iii) Mycoplasma hyorhinis (strain 38983), (iv) Mycoplasma hyosynoviae (strain 34428), and (v) M. hyopneumoniae (strain 232). Weekly serum and daily oral fluid samples were collected through 56 days postinoculation (dpi). The true status of pigs was established by PCR testing on oral fluids samples over the course of the observation period. Analysis of ELISA performance at various cutoffs found that the manufacturers’ recommended cutoffs were diagnostically specific, i.e., produced no false positives, with the exceptions of 2 ELISAs. An analysis based on overall misclassification error rates found that 4 ELISAs performed similarly, although one assay produced more false positives. In study 2, the 3 best-performing ELISAs from study 1 were compared using serum samples generated under field conditions. Ten 8-week-old pigs were intratracheally inoculated with M. hyopneumoniae. Matched serum and tracheal samples (to establish the true pig M. hyopneumoniae status) were collected at 7- to 14-day intervals through 98 dpi. Analyses of sensitivity and specificity showed similar performance among these 3 ELISAs. Overall, this study provides an assessment of the performance of current M. hyopneumoniae ELISAs and an understanding of their use in surveillance.

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