scholarly journals Diagnostic Techniques To Detect Cryptic Leishmaniasis in Dogs

2002 ◽  
Vol 9 (5) ◽  
pp. 1137-1141 ◽  
Author(s):  
Laura Iniesta ◽  
Salceda Fernández-Barredo ◽  
Béatrice Bulle ◽  
M. Teresa Gómez ◽  
Renaud Piarroux ◽  
...  
Keyword(s):  
Lymphocyte Proliferation ◽  
Antibody Test ◽  
Diagnostic Techniques ◽  
Delayed Type Hypersensitivity ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Proliferation Assay ◽  
Immunofluorescence Antibody Test ◽  
Immunofluorescence Antibody

ABSTRACT This study of several techniques for detecting cryptic leishmaniasis in dogs from areas in Spain where Leishmania infantum is highly endemic concludes that immunological techniques (enzyme-linked immunosorbent assay, immunofluorescence antibody test, Western blotting, delayed-type hypersensitivity reaction, and in vitro lymphocyte proliferation assay) do not clearly differentiate between noninfected and infected asymptomatic dogs and that culture and PCR are more reliable diagnostic tools.

scholarly journals Toward Diagnosing Leishmania infantum Infection in Asymptomatic Dogs in an Area Where Leishmaniasis Is Endemic

2009 ◽  
Vol 16 (3) ◽  
pp. 337-343 ◽  
Author(s):  
D. Otranto ◽  
P. Paradies ◽  
D. de Caprariis ◽  
D. Stanneck ◽  
G. Testini ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Clinical Signs ◽  
Antibody Test ◽  
Diagnostic Methods ◽  
Canine Leishmaniasis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Canine Monocytic Ehrlichiosis ◽  
Immunofluorescence Antibody Test ◽  
Immunofluorescence Antibody

ABSTRACT The most frequently used diagnostic methods were compared in a longitudinal survey with Leishmania infantum-infected asymptomatic dogs from an area of Italy where leishmaniasis is endemic. In February and March 2005, 845 asymptomatic dogs were tested by an immunofluorescence antibody test (IFAT), a dipstick assay (DS), and an enzyme-linked immunosorbent assay (ELISA) for L. infantum and by IFAT for Ehrlichia canis. Dogs seronegative for L. infantum were further parasitologically evaluated by microscopic examination of lymph node tissues and PCR of skin samples. A total of 204 animals both serologically and parasitologically negative for L. infantum at the first sampling were enrolled in the trial and were further examined for canine leishmaniasis (CanL) and canine monocytic ehrlichiosis in November 2005 (i.e., the end of the first sandfly season) and March 2006 and 2007 (1- and 2-year follow-ups, respectively). At the initial screening, the overall rates of L. infantum seroprevalence were 9.5% by IFAT, 17.1% by ELISA, and 9.8% by DS and the overall rate of E. canis seroprevalence was 15%. The rates of concordance between the results of IFAT and DS were almost equal, whereas the rate of concordance between the results of IFAT and DS and those of the ELISA was lower. The results of the annual incidence of Leishmania infection were variable, depending on the test employed, with the highest values registered for PCR (i.e., 5.7% and 11.4% at the 1- and 2-year follow-ups, respectively), followed by ELISA, IFAT, and DS. Over the 2 years of observation, 55 animals (i.e., 26.9%) became positive for L. infantum by one or more diagnostic tests at different follow-up times, with 12.7% showing clinical signs related to CanL, while the remaining 87.3% were asymptomatic. A diagnostic scheme for assessment of the L. infantum infection status in asymptomatic dogs is suggested.

scholarly journals Evaluation of the NovaLisa™ Leishmania Infantum IgG ELISA in A Reference Diagnostic Laboratory in A Non-Endemic Country

Antibodies ◽  
2019 ◽  
Vol 8 (1) ◽  
pp. 20 ◽  
Author(s):  
Christen Stensvold ◽  
Amalie Høst ◽  
Salem Belkessa ◽  
Henrik Nielsen
Keyword(s):  
Western Blot ◽  
Leishmania Infantum ◽  
Antibody Test ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Diagnostic Laboratory ◽  
Cost Effective Alternative ◽  
Immunofluorescence Antibody Test ◽  
Immunofluorescence Antibody

Anti-Leishmania antibodies may be detectable in patients with leishmaniasis. Here, we compared a commercial enzyme-linked immunosorbent assay (ELISA) for the detection of anti-Leishmania antibodies, with an immunofluorescence antibody test (IFAT) that is no longer commercially available. Eighty-six serum samples from 73 patients were tested. The results obtained by the NovaLisa™ Leishmania infantum IgG ELISA, interpreted according to the instructions of the manufacturer, but with a modified cut-off for borderline positive values, were compared with the IFAT results that were already available. Moreover, Leishmania Western blot IgG results were available for 43 of the samples. The overall concordance of ELISA and IFAT was 67%. The ELISA and IFAT tests scored as 24% and 15% of the samples being positive, respectively, while 13% and 33% scored as borderline-positive, respectively. Using a Western blot (WB) as the reference, the sensitivities and specificities for the positive plus borderline-positive samples combined was 95.5% (95% confidence interval (CI), 77.2–99.9%) and 81.0% (95% CI, 58.1–94.6%) for ELISA, and 95.5% (95% CI, 77.2–99.9%) and 42.9% (95% CI, 21.8–66.0%) for IFAT, respectively. Overall, the ELISA proved to be a cost-effective alternative to the IFAT, due to its higher accuracy and specificity, and with a consequently lower number of confirmatory WB tests being required. Lastly, we also present data on the associations between seroconversion and the type of leishmaniasis.

Trypanosoma congolense: appearance and distribution of variable antigen types during metacyclic differentiation in vitro

Parasitology ◽  
1990 ◽  
Vol 100 (1) ◽  
pp. 107-113 ◽  
Author(s):  
C. J. Prain ◽  
C. A. Ross
Keyword(s):  
Culture System ◽  
Antibody Test ◽  
Immunogold Labelling ◽  
Trypanosoma Congolense ◽  
Attachment Sites ◽  
Variable Antigen ◽  
Labelling Method ◽  
Immunofluorescence Antibody Test ◽  
Immunofluorescence Antibody

SummaryDifferentiation of epimastigotes and production of infective metacyclic forms of Trypanosoma congolense were examined in a culture system which enabled manipulation of the population density of insect forms. Scanning electron microscopy of cultures revealed the attachment sites of epimastigotes in detail, showing them to be attached as ‘clusters’ or ‘bundles’ and having associated fibrillar structures. Dividing epimastigotes were observed either within individual bundles or in association with two bundles. Metacyclic forms were detected by an immunofluorescence antibody test (IFAT) using metacyclic variable-antigen type (M-VAT) specific monoclonal antibodies, by day 7 after seeding cultures. Trypanosomes expressing M-VATs appeared singly in bundles, observed by both IFAT and an immunogold labelling method. Statistical analysis using Poisson calculations suggested that, in general, the distribution of metacyclics expressing individual M-VATs was random throughout cultures.

scholarly journals Immune Responses to Defined Antigens of Mycobacterium bovis in Cattle Experimentally Infected with Mycobacterium kansasii

2006 ◽  
Vol 13 (6) ◽  
pp. 611-619 ◽  
Author(s):  
W. R. Waters ◽  
M. V. Palmer ◽  
T. C. Thacker ◽  
J. B. Payeur ◽  
N. B. Harris ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Bovine Tuberculosis ◽  
Nontuberculous Mycobacteria ◽  
Gamma Interferon ◽  
Delayed Type Hypersensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Antibodies ◽  
Specific Serum ◽  
Specific Proteins

ABSTRACT Cross-reactive responses elicited by exposure to nontuberculous mycobacteria often confound the interpretation of antemortem tests for Mycobacterium bovis infection of cattle. The use of specific proteins (e.g., ESAT-6, CFP-10, and MPB83), however, generally enhances the specificity of bovine tuberculosis tests. While genes for these proteins are absent from many nontuberculous mycobacteria, they are present in M. kansasii. Instillation of M. kansasii into the tonsillar crypts of calves elicited delayed-type hypersensitivity and in vitro gamma interferon and nitrite concentration responses of leukocytes to M. avium and M. bovis purified protein derivatives (PPDs). While the responses of M. kansasii-inoculated calves to M. avium and M. bovis PPDs were approximately equivalent, the responses of M. bovis-inoculated calves to M. bovis PPD exceeded their respective responses to M. avium PPD. The gamma interferon and nitrite responses of M. kansasii-inoculated calves to recombinant ESAT-6-CFP-10 (rESAT-6-CFP-10) exceeded corresponding responses of noninoculated calves as early as 15 and 30 days after inoculation, respectively, and persisted throughout the study. The gamma interferon and nitrite responses of M. bovis-inoculated calves to rESAT-6-CFP-10 exceeded the corresponding responses of M. kansasii-inoculated calves beginning 30 days after inoculation. By using a lipoarabinomannan-based enzyme-linked immunosorbent assay, specific serum antibodies were detected as early as 50 days after challenge with M. kansasii. By a multiantigen print immunoassay and immunoblotting, serum antibodies to MPB83, but not ESAT-6 or CFP-10, were detected in M. kansasii-inoculated calves; however, responses to MPB83 were notably weaker than those elicited by M. bovis infection. These findings indicate that M. kansasii infection of calves elicits specific responses that may confound the interpretation of bovine tuberculosis tests.

scholarly journals Seroprevalence of Toxoplasma gondii in dogs of riverside communities of Mato Grosso Pantanal, Brazil

2016 ◽  
Vol 25 (4) ◽  
pp. 531-535 ◽  
Author(s):  
Juliana Yuki Rodrigues ◽  
Arleana do Bom Parto Ferreira de Almeida ◽  
Eveline da Cruz Boa Sorte ◽  
Naiani Domingos Gasparetto ◽  
Felipe Augusto Constantino Seabra da Cruz ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Antibody Test ◽  
Human Infection ◽  
Mato Grosso ◽  
Exact Test ◽  
Worldwide Distribution ◽  
Significant Difference ◽  
Chi Squared ◽  
Immunofluorescence Antibody Test ◽  
Immunofluorescence Antibody

Abstract Toxoplasma gondii is an intracellular protozoan with worldwide distribution and dogs act as sentinels of human infection. This search aimed to determine the occurrence of antibodies against T. gondii in dogs of the communities on the Cuiabá River, Mato Grosso and variables associated with infection. The dogs of the riverside communities in Cuiabá River, which includes Barranco Alto, Praia do Poço, Engenho Velho, Varginha, Bom Sucesso, Passagem da Conceição and São Gonçalo Beira Rio, were evaluated for the presence of T. gondii antibodies by indirect immunofluorescence antibody test (IFAT). The prevalence and factors associated with infection were calculated by chi-squared test (χ2) or Fisher’s exact test, and univariate and multiple analysis. Of the 248 dogs surveyed, 107 (43.1%) were seropositive for T. gondii. The seroprevalence ranged from 25.6% to 64.3%. There was no statistically significant difference between the communities studied (p > 0.05). As for the associated factors, the only statistically significant factor was that of dogs living with cats (p = 0.02), with approximately twice the risk of acquiring infection. In conclusion, the seroprevalence in dogs of riverside communities in the Baixada Cuiabana demonstrated that high rates of infection, being the factor associated with infection, contact with domestic cats.

scholarly journals Safety of measles, mumps, and rubella vaccine in egg allergy:in vivo and in vitro management

2020 ◽  
Author(s):  
Stefania Magistà ◽  
Marcello Albanesi ◽  
Nada Chaoul ◽  
Danilo Di Bona ◽  
Elisabetta Di Leo ◽  
...  
Keyword(s):  
Lymphocyte Proliferation ◽  
B Lymphocyte ◽  
Ex Vivo ◽  
Diagnostic Strategy ◽  
Egg Allergy ◽  
Lymphocyte Proliferation Assay ◽  
Proliferation Assay ◽  
Mmr Vaccine

Abstract Background Egg allergy is the second most prevalent form of food allergy in childhood. In spite of the evidence accumulated, inoculating egg allergy children with attenuated vaccines grown on chick embryo cell cultures, such as the measles, mumps, and rubella (MMR) vaccine, is regarded (erroneously) as potentially dangerous or even anaphylactogenic, by many. An issue perceived as particularly conflicting also by Health Professionals.Case presentation A 15-year-old boy, with a history of severe egg allergy in early infancy, who was still sensitized to egg allergens, including baked egg, had never received MMR vaccination, in fear of possible anaphylaxis, in spite of the fact that this vaccination is mandatory in the first year of life, in Italy. Because of that, he was not allowed to attend school, longer, and was referred to us in order to assess the potential risk of MMR vaccination. Upon thorough allergologic workup, sensitization to MMR vaccine components was excluded by an in vivo approach, consisting in skin prick tests, intradermal tests, and subcutaneous injection test, corroborated by vaccine-specific B-lymphocyte proliferation assay, ex vivo. T-cell proliferation in response to MMR vaccine was also excluded. Eventually, the boy was inoculated with MMR vaccine and was readmitted to school.Conclusions The diagnostic strategy adopted appears feasible and easy-to-perform and may be adopted in controversial cases (as the one reported), characterized by previous severe allergic reactions to egg. The B-lymphocyte proliferation assay we developed may represent a useful and reliable tool not only in research but also in clinical practice.

scholarly journals Application of Volatilome Analysis to the Diagnosis of Mycobacteria Infection in Livestock

2021 ◽  
Vol 8 ◽  
Author(s):  
Pablo Rodríguez-Hernández ◽  
Vicente Rodríguez-Estévez ◽  
Lourdes Arce ◽  
Jaime Gómez-Laguna
Keyword(s):  
Analytical Techniques ◽  
Diagnostic Techniques ◽  
Diagnostic Tools ◽  
Future Research ◽  
Control Programs ◽  
Targeted Analysis ◽  
Low Sensitivity ◽  
And Control

Volatile organic compounds (VOCs) are small molecular mass metabolites which compose the volatilome, whose analysis has been widely employed in different areas. This innovative approach has emerged in research as a diagnostic alternative to different diseases in human and veterinary medicine, which still present constraints regarding analytical and diagnostic sensitivity. Such is the case of the infection by mycobacteria responsible for tuberculosis and paratuberculosis in livestock. Although eradication and control programs have been partly managed with success in many countries worldwide, the often low sensitivity of the current diagnostic techniques against Mycobacterium bovis (as well as other mycobacteria from Mycobacterium tuberculosis complex) and Mycobacterium avium subsp. paratuberculosis together with other hurdles such as low mycobacteria loads in samples, a tedious process of microbiological culture, inhibition by many variables, or intermittent shedding of the mycobacteria highlight the importance of evaluating new techniques that open different options and complement the diagnostic paradigm. In this sense, volatilome analysis stands as a potential option because it fulfills part of the mycobacterial diagnosis requirements. The aim of the present review is to compile the information related to the diagnosis of tuberculosis and paratuberculosis in livestock through the analysis of VOCs by using different biological matrices. The analytical techniques used for the evaluation of VOCs are discussed focusing on the advantages and drawbacks offered compared with the routine diagnostic tools. In addition, the differences described in the literature among in vivo and in vitro assays, natural and experimental infections, and the use of specific VOCs (targeted analysis) and complete VOC pattern (non-targeted analysis) are highlighted. This review emphasizes how this methodology could be useful in the problematic diagnosis of tuberculosis and paratuberculosis in livestock and poses challenges to be addressed in future research.

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