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VETERINARY IMMUNOLOGY

Isolation of Potentially Useful Antigens from Cyathostomin Third-Stage Larvae by Using a Fast Protein Liquid Chromatography One-Step Method

A. Paz-Silva, R. Francisco, I. Rodríguez, I. Francisco, C. F. Cazapal-Monteiro, M. S. Arias, J. L. Suárez, R. Sánchez-Andrade
A. Paz-Silva
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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  • For correspondence: adolfo.paz@usc.es
R. Francisco
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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I. Rodríguez
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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I. Francisco
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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C. F. Cazapal-Monteiro
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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M. S. Arias
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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J. L. Suárez
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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R. Sánchez-Andrade
Equine Diseases Study Group (Epidemiology, Parasitology and Zoonoses), Animal Pathology Department, Veterinary Faculty, Santiago de Compostela University, 27002-Lugo, Spain
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DOI: 10.1128/CVI.05189-11
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  • Fig. 1.
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    Fig. 1.

    Chromatogram of the FPLC protein complexes isolated from a mixture of cyathostomin third-stage larvae. AU, arbitrary units.

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    Fig. 2.

    ROC analysis of the results achieved by using the FPLC protein complexes isolated from a mixture of cyathostomin third-stage larvae and sera from infected and uninfected horses.

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    Fig. 3.

    Kinetics of cyathostomin egg output in naturally infected weanlings. Values along the y axis represent numbers of cythostomin eggs per gram of feces (EPG). Data represent the mean values plus 2 standard deviations.

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    Fig. 4.

    Kinetics of IgG(T) antibodies against antigenic peaks P1 (A), P2 (B), and P3 (C) isolated from L3 cyathostomin excretory/secretory antigens by means of an FPLC system. Values along the y axis represent optical density at 492 nm. Data represent the mean values plus 2 standard deviations.

Tables

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  • Table 1.

    Gel filtration chromatography protocol

    StepVol (ml)Description
    10.00Collection of 4-ml fractions during entire run
    20.00Lamp (UV detector) turned on
    30.00Zero baseline
    40.00Isocratic flow
    55.00Load/inject 2.5-ml sample
    67.5Isocratic flow
    7600End of protocol
  • Table 2.

    Analysis of the liquid chromatography (FPLC) protein complexes isolated from a mixture of cyathostomin third-stage larvaea

    Antigen peak and size (kDa)Cutoff valueAUCPS (%)SP (%)PLRNLR
    P1 (51)0.48500.9820.0019298460.08
    P2 (29)0.49650.9850.0019294150.08
    P3 (15)0.48150.9820.0019291100.09
    • ↵a The cutoff point for each antigen was established on the basis of the highest values achieved for the AUC (maximum 1), S (sensitivity, 0 to 100), SP (specificity, 0 to 100), PLR (positive-likelihood ratio; elevated numbers are expected), and NLR (negative-likelihood ratio; low values are expected).

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Isolation of Potentially Useful Antigens from Cyathostomin Third-Stage Larvae by Using a Fast Protein Liquid Chromatography One-Step Method
A. Paz-Silva, R. Francisco, I. Rodríguez, I. Francisco, C. F. Cazapal-Monteiro, M. S. Arias, J. L. Suárez, R. Sánchez-Andrade
Clinical and Vaccine Immunology Aug 2011, 18 (9) 1462-1466; DOI: 10.1128/CVI.05189-11

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Isolation of Potentially Useful Antigens from Cyathostomin Third-Stage Larvae by Using a Fast Protein Liquid Chromatography One-Step Method
A. Paz-Silva, R. Francisco, I. Rodríguez, I. Francisco, C. F. Cazapal-Monteiro, M. S. Arias, J. L. Suárez, R. Sánchez-Andrade
Clinical and Vaccine Immunology Aug 2011, 18 (9) 1462-1466; DOI: 10.1128/CVI.05189-11
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