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Serodiagnosis of fasciolosis by fast protein liquid chromatography-fractionated excretory/secretory antigens

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Abstract

In several studies, different antigenic preparations and diverse immunological tests were applied for serodiagnosis of Fasciola hepatica infections. Most of these preparations showed cross-reactivity with proteins of other parasites. Application of purified antigens might reduce these cross-reactivities. Here, we used fast protein liquid chromatography (FPLC)-fractionated extracts of F. hepatica excretory/secretory antigens (E/S Ags) for serodiagnosis of human and sheep fasciolosis. To develop an improved diagnostic method, we fractionated F. hepatica E/S Ags by anion exchange chromatography on a Sepharose CL-6B column and then tested the serodiagnostic values of the fractions. We used sera from F. hepatica-infected human and sheep as positive controls. Sera from patients with hydatidosis and strongyloidiasis were used for cross-reactivity studies. Enzyme-linked immunosorbent assays (ELISA) of the second FPLC peak, containing 20, 25, and 70 kDa proteins, discriminated between F. hepatica-infected and uninfected human and sheep samples. Fractionation of F. hepatica E/S Ags by FPLC is a fast and reproducible way of obtaining antigens useful for serodiagnosis of human and sheep fasciolosis with acceptable sensitivity and specificity.

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Acknowledgments

This study was supported by grant 24488 from the Iran University of Medical Sciences (IUMS), Tehran, Iran. We thank Professor Eshrat beigom Kia from the School of Public Health, Tehran University of Medical Sciences (Tehran, Iran) for providing strongyloidiasis sera and Professor AbdolFatah Sarafinejhad from the Noor Referral Laboratory (Tehran, Iran) for providing us with fasciolosis patients’ sera.

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Authors and Affiliations

  1. Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences (IUMS), Tehran, Iran

    Kobra Mokhtarian, Lame Akhlaghi, Ahmad Reza Meamar & Elham Razmjou

  2. Department of Parasitology and Mycology, School of Medicine, Shahrekord University of Medical Science, Shahrekord, Iran

    Kourosh Manouchehri Naeini

  3. Immunology Research Center, Iran University of Medical Science (IUMS), Tehran, Iran

    Samaneh Gholami, Masoomeh Najafi Samei & Reza Falak

  4. Department of Immunology, School of Medicine, Iran University of Medical Sciences (IUMS), Tehran, Iran

    Masoomeh Najafi Samei & Reza Falak

Authors
  1. Kobra Mokhtarian
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  2. Lame Akhlaghi
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  3. Ahmad Reza Meamar
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  4. Elham Razmjou
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  5. Kourosh Manouchehri Naeini
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  6. Samaneh Gholami
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  7. Masoomeh Najafi Samei
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  8. Reza Falak
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Corresponding authors

Correspondence to Lame Akhlaghi or Reza Falak.

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Highlights

• Fasciola hepatica antigens were fractionated by FPLC anion exchange chromatography.

• Fractions containing the 17-, 48-, and 50-kDa proteins cross-reacted with sera from hydatidosis patients.

• The 12-, 14-, and 25-kDa proteins discriminate fasciolosis with high sensitivity and specificity.

• FPLC is a fast, simple, and reproducible way to purify F. hepatica immunogens.

• FPLC-fractionated antigens are useful for serodiagnosis of human and sheep fasciolosis.

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Mokhtarian, K., Akhlaghi, L., Meamar, A.R. et al. Serodiagnosis of fasciolosis by fast protein liquid chromatography-fractionated excretory/secretory antigens. Parasitol Res 115, 2957–2965 (2016). https://doi.org/10.1007/s00436-016-5049-7

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  • DOI: https://doi.org/10.1007/s00436-016-5049-7

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