Identification and Preliminary Evaluation of a Novel Recombinant Protein for Serodiagnosis of Strongyloidiasis

Norsyahida Arifin Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia;

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Muhammad Hafiznur Yunus Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia;

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Thomas J. Nolan Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania

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James B. Lok Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania

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Rahmah Noordin Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia;

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Strongyloides stercoralis is a human parasite that can cause a long-term infection. In immunosuppressed patients, strongyloidiasis may be fatal when there is overwhelming autoinfection resulting in the migration of large numbers of larvae through many organs. Definitive diagnosis is still a challenge, and a combination of symptoms, microscopic identification, and serology test results are often used to arrive at a clinical decision. However, intermittent larval excretion, low parasite burden, and occult infections are challenges with parasitological diagnosis of infection with S. stercoralis. Meanwhile, serologic tests using immunoglobulin G and parasite antigen extract have problems of cross-reactivity with other helminthic infections. Recombinant antigen-based serodiagnosis is a good alternative to overcome the laboratory diagnostic issues. Herein, we report on the isolation of cDNA clone encoding an antigen of potential diagnostic value identified from immunoscreening of a S. stercoralis cDNA library. The translated protein had highest similarity to Strongyloides ratti immunoglobulin-binding protein 1. The recombinant antigen produced, rSs1a, was assessed using western blot and enzyme-linked immunosorbent assay. The latter showed 96% diagnostic sensitivity and 93% specificity; thus, rSs1a has good potential for use in serodiagnosis of human strongyloidiasis.

Author Notes

Address correspondence to Rahmah Noordin, Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800 Penang, Malaysia. E-mail: rahmah8485@gmail.com

Financial support: Major funding for this project was provided by the Malaysian Ministry of Higher Education (ERGS 203/CIPPM/6730048, HICoE 311/CIPPM/4401005) and Universiti Sains Malaysia (USM-RU 1001/CIPPM/812078). The National Institutes of Health (NIH) grants AI105856 and AI22662 to JBL and NIH Referral Center grant OD P40-10939 to Dr. Charles Vite provided research materials for this project.

Authors’ addresses: Norsyahida Arifin, Muhammad Hafiznur Yunus, and Rahmah Noordin, Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800 Penang, Malaysia, E-mails: syahidaarifin@gmail.com, mhn1016@yahoo.com, and rahmah8485@gmail.com. Thomas J. Nolan and James B. Lok, Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, E-mails: parasit@vet.upenn.edu and jlok@vet.upenn.edu.

  • 1.

    Bisoffi Z et al. 2013. Strongyloides stercoralis: a plea for action. PLoS Negl Trop Dis 7: e2214.

  • 2.

    Angheben A, Mistretta M, Gobbo M, Bonafini S, Iacovazzi T, Sepe A, Gobbi F, Marocco S, Rossanese A & Bisoffi Z 2011. Acute strongyloidiasis in Italian tourists returning from southeast Asia. J Travel Med 18: 138140.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 3.

    Keiser PB & Nutman TB 2004. Strongyloides stercoralis in the immunocompromised population. Clin Microbiol Rev 17: 208217.

  • 4.

    Simon GL 2009. Strongyloidiasis. Satoskar AR, Simon GL, Hotez PJ, Tsuji M, eds. Medical Parasitology. Austin, TX: Landes Bioscience, 31–38.

    • PubMed
    • Export Citation
  • 5.

    Olsen A, van Lieshout L, Marti H, Polderman T, Polman K, Steinmann P, Stothard R, Thybo S, Verweij JJ & Magnussen P 2009. Strongyloidiasis—the most neglected of the neglected tropical diseases? Trans R Soc Trop Med Hyg 103: 967972.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6.

    Lindo JF & Lee MG 2001. Strongyloides stercoralis and S. fulleborni. Gillespie SH, Pearson RD, eds. Principles and Practice of Clinical Parasitology. New York, NY: Wiley Online Library, 479–492.

    • PubMed
    • Export Citation
  • 7.

    Prendki V, Fenaux P, Durand R, Thellier M, Bouchaud O, 2011. Strongyloidiasis in man 75 years after initial exposure. Emerg Infect Dis 17: 931932.

  • 8.

    Siddiqui AA, Stanley CS & Berk SL 2001. Isolation of a cDNA encoding an IgG immunoreactive antigen (zinc finger protein) of Strongyloides stercoralis. Parasite 8: 163167.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9.

    Iriemenam NC, Sanyaolu AO, Oyibo WA & Fagbenro-Beyioku AF 2010. Strongyloides stercoralis and the immune response. Parasitol Int 59: 914.

  • 10.

    Ramachandran S, Thompson RW, Gam AA & Neva FA 1998. Recombinant cDNA clones for immunodiagnosis of strongyloidiasis. J Infect Dis 177: 196203.

  • 11.

    Ravi V, Ramachandran S, Thompson RW, Andersen JF & Neva FA 2002. Characterization of a recombinant immunodiagnostic antigen (NIE) from Strongyloides stercoralis L3-stage larvae. Mol Biochem Parasitol 125: 7381.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Anderson NW, Klein DM, Dornink SM, Jespersen DJ, Kubofcik J, Nutman TB, Merrigan SD, Couturier MR & Theel ES 2014. Comparison of three immunoassays for detection of antibodies to Strongyloides stercoralis. Clin Vaccine Immunol 21: 732736.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Norsyahida A, Riazi M, Sadjjadi SM, Muhammad Hafiznur Y, Low HC, Zeehaida M & Noordin R 2013. Laboratory detection of strongyloidiasis: IgG-, IgG4 - and IgE-ELISAs and cross-reactivity with lymphatic filariasis. Parasite Immunol 35: 174179.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14.

    Amerizadeh A, Idris ZM, Khoo BY, Kotresha D, Yunus MH, Karim IZ, Saadatnia G, Teh AY & Noordin R 2013. Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera. Microb Pathog 54: 6066.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15.

    Kobayashi Y, Ohsaki K, Ikeda K, Kakemoto S, Ishizaki S, Shimakura K, Nagashima Y & Shiomi K 2011. Identification of novel three allergens from Anisakis simplex by chemiluminescent immunoscreening of an expression cDNA library. Parasitol Int 60: 144150.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16.

    Wu XP, Fu BQ, Wang XL, Yu L, Yu SY, Deng HK, Liu XY, Boireau P, Wang F & Liu MY 2009. Identification of antigenic genes in Trichinella spiralis by immunoscreening of cDNA libraries. Vet Parasitol 159: 272275.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17.

    Li J, Zhang WB & McManus DP 2004. Recombinant antigens for immunodiagnosis of cystic echinococcosis. Biol Proced Online 6: 6777.

  • 18.

    Siddiqui AA, Stanley CS & Berk SL 2000. A cDNA encoding the highly immunodominant antigen of Strongyloides stercoralis: gamma-subunit of isocitrate dehydrogenase (NAD+). Parasitol Res 86: 279283.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19.

    Siddiqui AA, Stanley CS, Skelly PJ & Berk SL 2000. A cDNA encoding a nuclear hormone receptor of the steroid/thyroid hormone-receptor superfamily from the human parasitic nematode Strongyloides stercoralis. Parasitol Res 86: 2429.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20.

    Gallego SG, Slade RW & Brindley PJ 1998. A cDNA encoding a pepsinogen-like, aspartic protease from the human roundworm parasite Strongyloides stercoralis. Acta Trop 71: 1726.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21.

    Ramanathan R, Burbelo PD, Groot S, Iadarola MJ, Neva FA & Nutman TB 2008. A luciferase immunoprecipitation systems assay enhances the sensitivity and specificity of diagnosis of Strongyloides stercoralis infection. J Infect Dis 198: 444451.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22.

    Rascoe LN, Price C, Shin SH, McAuliffe I, Priest JW & Handali S 2015. Development of Ss-NIE-1 recombinant antigen based assays for immunodiagnosis of strongyloidiasis. PLoS Negl Trop Dis 9: e0003694.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23.

    Atkins NS, Conway DJ, Lindo JF, Bailey JW & Bundy DA 1999. L3 antigen-specific antibody isotype responses in human strongyloidiasis: correlations with larval output. Parasite Immunol 21: 517526.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24.

    Genta RM, Schad GA & Hellman ME 1986. Strongyloides stercoralis: parasitological, immunological and pathological observations in immunosuppressed dogs. Trans R Soc Trop Med Hyg 80: 3441.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25.

    Schaffel R, Nucci M, Carvalho E, Braga M, Almeida L, Portugal R & Pulcheri W 2001. The value of an immunoenzymatic test (enzyme-linked immunosorbent assay) for the diagnosis of strongyloidiasis in patients immunosuppressed by hematologic malignancies. Am J Trop Med Hyg 65: 346350.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 26.

    Conway DJ, Bailey JW, Lindo JF, Robinson RD, Bundy DA & Bianco AE 1993. Serum IgG reactivity with 41-, 31-, and 28-kDa larval proteins of Strongyloides stercoralis in individuals with strongyloidiasis. J Infect Dis 168: 784787.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 27.

    Conway DJ, Lindo JF, Robinson RD, Bundy DA & Bianco AE 1994. Strongyloides stercoralis: characterization of immunodiagnostic larval antigens. Exp Parasitol 79: 99105.

  • 28.

    Atkins NS, Lindo JF, Lee MG, Conway DJ, Bailey JW, Robinson RD & Bundy DA 1997. Humoral responses in human strongyloidiasis: correlations with infection chronicity. Trans R Soc Trop Med Hyg 91: 609613.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 29.

    Rodrigues RM, de Oliveira MC, Sopelete MC, Silva DA, Campos DM, Taketomi EA & Costa-Cruz JM 2007. IgG1, IgG4, and IgE antibody responses in human strongyloidiasis by ELISA using Strongyloides ratti saline extract as heterologous antigen. Parasitol Res 101: 12091214.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 30.

    Satoh M, Toma H, Sato Y, Kikuchi M, Takara M, Shiroma Y, Kiyuna S & Hirayama K 1999. Production of a high level of specific IgG4 antibody associated with resistance to albendazole treatment in HLA-DRB1*0901-positive patients with strongyloidiasis. Am J Trop Med Hyg 61: 668671.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31.

    Shields AM, Thompson SJ, Panayi GS & Corrigall VM 2012. Pro-resolution immunological networks: binding immunoglobulin protein and other resolution-associated molecular patterns. Rheumatology (Oxford) 51: 780788.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 32.

    Morris JA, Dorner AJ, Edwards CA, Hendershot LM & Kaufman RJ 1997. Immunoglobulin binding protein (BiP) function is required to protect cells from endoplasmic reticulum stress but is not required for the secretion of selective proteins. J Biol Chem 272: 43274334.

    • PubMed
    • Search Google Scholar
    • Export Citation
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