Skip to main content
SpringerLink
Log in

Multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis

  • Original Paper
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Antigenic proteins from Clonorchis sinensis have been previously purified and evaluated for their antigenicity to enable the serodiagnosis of clonorchiasis. Though they were of high specificity, molecularly defined proteins were reported to be less sensitive as single antigens than crude antigen. To resolve this issue, 11 clones were selected by immunoscreening an adult C. sinensis cDNA library using infected human sera. Mixed antigens were prepared using recombinant proteins of positive clones and investigated for antigenicity by immunoblotting against C. sinensis- and helminth-infected patient sera. A mixed antigen of recombinant 28 and 26 kDa glutathion S-transferases (Cs28GST and Cs26GST) produced 76% sensitivity and 95% specificity. Furthermore, a triple mix of recombinant Cs26GST and Cs28GST with vitelline precursor protein pushed up the sensitivity to 87% and maintained specificity at 95%. It is proposed that multiple antigen mixes should be further studied to develop rapid serodiagnostic test kits for the serodiagnosis of human clonorchiasis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed  CAS  Google Scholar 

  • Bouvard V, Baan R, Straif K, Grosse Y, Secretan B, El Ghissassi F, Benbrahim-Tallaa L, Guha N, Freeman C, Galichet L, Cogliano V, WHO International Agency for Research on Cancer Monograph Working Group (2009) A review of human carcinogens-Part B: biological agents. Lancet Oncol 10:321–322

    Article  PubMed  Google Scholar 

  • Charest H, Matlashewski G (1994) Developmental gene expression in Leishmania donovani: differential cloning and analysis of an amastigote-stage-specific gene. Mol Cell Biol 14:2975–2984

    PubMed  CAS  Google Scholar 

  • Choi MH, Park IC, Li S, Hong ST (2003) Excretory-secretory antigen is better than crude antigen for the serodiagnosis of clonorchiasis by ELISA. Korean J Parasitol 41:35–39

    Article  PubMed  Google Scholar 

  • Eursitthichai V, Viyanant V, Tesana S, Sithithawon P, Kosa N, Grams R (2010) Oipsthorchis viverrini: Evaluation of 28 kDa glutathione S-transferase as diagnostic tool in human opisthorchiasis. Acta Trop 114:76–80

    Article  PubMed  CAS  Google Scholar 

  • Hong ST (1988) Changes of anti-Clonorchis sinensis IgG antibody in serum after praziquantel treatment in human clonorchiasis. Korean J Parasitol 26:1–8

    Article  Google Scholar 

  • Hong ST, Kho WG, Lee M, Lee JS, Lee SH (1997) Immunoblot patterns of clonorchiasis. Korean J Parasitol 35:87–93

    Article  PubMed  CAS  Google Scholar 

  • Hong SJ, Seong KY, Sohn WM, Song KY (2000) Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis. Mol Biochem Parasitol 108:207–216

    Article  PubMed  CAS  Google Scholar 

  • Hong SJ, Lee JY, Lee DH, Sohn WM, Cho SY (2001) Molecular cloning and characterization of a mu-class glutathione S-transferase from Clonorchis sinensis. Mol Biochem Parasitol 115:69–75

    Article  PubMed  CAS  Google Scholar 

  • Hong SJ, Yun Kim T, Gan XX, Shen LY, Sukontason K, Sukontason K, Kang SY (2002) Clonorchis sinensis: glutathione S-transferase as a serodiagnostic antigen for detecting IgG and IgE antibodies. Exp Parasitol 101:231–233

    Article  PubMed  CAS  Google Scholar 

  • Hu F, Yu X, Ma C, Zhou H, Zhou Z, Li Y, Lu F, Xu J, Wu Z, Hu X (2007) Clonorchis sinensis: Expression, characterization, immunolocalization and serological reactivity of one excretory/secretory antigen-LPAP homologue. Exp Parasitol 117:157–164

    Article  PubMed  CAS  Google Scholar 

  • Imboden M, Müller N, Hemphill A, Mattioli R, Seebeck T (1995) Repetitive proteins from the flagellar cytoskeleton of African trypanosomes are diagnostically useful antigens. Parasitology 110:249–258

    Article  PubMed  CAS  Google Scholar 

  • Kang SY, Ahn IY, Park CY, Chung YB, Hong ST, Kong Y, Cho SY, Hong SJ (2001) Clonorchis sinensis: molecular cloning and characterization of 28-kDa glutathione S-transferase. Exp Parasitol 97:186–195

    Article  PubMed  CAS  Google Scholar 

  • Kim TY, Kang SY, Ahn IY, Cho SY, Hong SJ (2001) Molecular cloning and characterization of an antigenic protein with a repeating region from Clonorchis sinensis. Korean J Parasitol 39:57–66

    Article  PubMed  CAS  Google Scholar 

  • Kim TY, Cho PY, Na JW, Hong SJ (2007) Molecular cloning and phylogenetic analysis of Clonorchis sinensis elongation factor-1α. Parasitol Res 101:1557–1562

    Article  PubMed  Google Scholar 

  • Kim TS, Cho SH, Huh S, Kong Y, Sohn WM, Hwang SS, Chai JY et al (2009) A nationwide survey on the prevalence of intestinal parasitic infections in the Republic of Korea, 2004. Korean J Parasitol 47:37–47

    Article  PubMed  Google Scholar 

  • Kwon YD, Cho PY, Hong SJ (2005) Clonorchis sinensis: molecular cloning and localization of myosin regulatory light chain. Parasitol Res 97:21–26

    Article  PubMed  Google Scholar 

  • Lee JH, Rim HJ, Bak UB (1993) Effect of Clonorchis sinensis infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian golden hamsters. Korean J Parasitol 31:21–30

    Article  PubMed  CAS  Google Scholar 

  • Lee KW, Shalaby KA, Thakur A, Medhat AM, Karim AM, LoVerde PT (1995a) Cloning of the gene for phosphoglycerate kinase from Schistosoma mansoni and characterization of its gene product. Mol Biochem Parasitol 71:221–231

    Article  PubMed  CAS  Google Scholar 

  • Lee KW, Thakur A, Karim AM (1995b) LoVerde PT (1995b) Immune response to Schistosoma mansoni phosphoglycerate kinase during natural and experimental infection: identification of a schistosome-specific B-cell epitope. Infect Immun 63:4307–4311

    PubMed  CAS  Google Scholar 

  • Li S, Chung BS, Choi MH, Hong ST (2004) Organ-specific antigens of Clonorchis sinensis. Korean J Parasitol 42:169–174

    Article  PubMed  Google Scholar 

  • Lim MK, Ju YH, Franceschi S, Oh JK, Kong HJ, Hwang SS, Park SK, Cho SI, Sohn WM, Kim DI, Yoo KY, Hong ST, Shin HR (2006) Clonorchis sinensis infection and increasing risk of cholangiocarcinoma in the Republic of Korea. Am J Trop Med Hyg 75:93–96

    PubMed  Google Scholar 

  • Lun ZR, Gasser RB, Lai DH, Li AX, Zhu XQ, Yu XB, Fang YY (2005) Clonorchiasis: a key food-borne zoonosis in China. Lancet Infect Dis 5:31–41

    Article  PubMed  Google Scholar 

  • Marín MS, Prieto M, Martín JM, Casais R, Boga JA, Parra F (1992) Identification and expression of a Fasciola hepatica gene encoding a gut antigen protein bearing repetitive sequences. Mol Biochem Parasitol 55:155–165

    Article  PubMed  Google Scholar 

  • Na BK, Lee HJ, Cho SH, Lee HW, Cho JH, Kho WG, Lee JS, Lee JS, Song KJ, Park PH, Song CY, Kim TS (2002) Expression of cysteine proteinase of Clonorchis sinensis and its use in serodiagnosis of clonorchiasis. J Parasitol 88:1000–1006

    PubMed  CAS  Google Scholar 

  • Nagano I, Pei F, Wu Z, Wu J, Cui H, Boonmars T, Takahashi Y (2004) Molecular expression of a cysteine proteinase of Clonorchis sinensis and its application to an enzyme-linked immunosorbent assay for immunodiagnosis of clonorchiasis. Clin Diagn Lab Immunol 11:411–416

    PubMed  CAS  Google Scholar 

  • Rim HJ (1986) The current pathobiology and chemotherapy of clonorchiasis. Korean J Parasitol 24:1–141

    Article  Google Scholar 

  • Rim HJ (2005) Clonorchiasis: an update. J Helminthol 79:269–281

    Article  PubMed  Google Scholar 

  • Shen C, Lee JA, Allam SRA, Bae YM, Han ET, Takeo S, Tsuboi T, Hong ST, Choi MH (2009) Serodiagnostic applicability of recombinant antigens of Clonorchis sinensis expressed by wheat germ cell-free protein synthesis system. Diagn Microbiol Inf Dis 64:334–339

    Article  CAS  Google Scholar 

  • Song CY, Dresden MH, Rege AA (1990) Clonorchis sinensis: purification and characterization of a cysteine proteinase from adult worms. Comp Biochem Physiol B 97:825–829

    Article  PubMed  CAS  Google Scholar 

  • Tang Y, Cho PY, Kim BS, Hong SJ (2005) Molecular cloning and characterization of vitelline precursor protein B from Clonorchis sinensis. J Parasitol 91:1374–1378

    Article  PubMed  CAS  Google Scholar 

  • Tang Y, Cho PY, Kim TI, Hong SJ (2006) Clonorchis sinensis: molecular cloning, enzymatic activity, and localization of yolk ferritin. J Parasitol 92:1275–1280

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  • Trottein F, Godin C, Pierce RJ, Sellin B, Taylor MG, Gorillot I, Silva MS, Lecocq JP, Capron A (1992) Inter-species variation of schistosome 28-kDa glutathione S-transferases. Mol Biochem Parasitol 54:63–72

    Article  PubMed  CAS  Google Scholar 

  • Yong TS, Yang HJ, Park SJ, Kim YK, Lee DH, Lee SM (1998) Immunodiagnosis of clonorchiasis using a recombinant antigen. Korean J Parasitol 36:183–190

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Jin Z, Da R, Dong Y, Song W, Chen X, Huang Q, Ling H, Che Y, Li Y, Zhang F (2008) Fas/FasL-dependent apoptosis of hepatocytes induced in rat and patients with Clonorchis sinensis infection. Parasitol Res 103:393–399

    Article  PubMed  Google Scholar 

  • Zhao QP, Moon SU, Lee HW, Na BK, Cho SY, Kong Y, Jiang MS, Li AH, Kim TS (2004) Evaluation of Clonorchis sinensis recombinant 7-kilodalton antigen for serodiagnosis of clonorchiasis. Clin Diagn Lab Immunol 11:814–817

    PubMed  CAS  Google Scholar 

Download references

Acknowledgement

This study was supported by the Ministry of Health and Welfare, Republic of Korea (Grant no. 01-PJ1-PG3-20200-0031). We extend our thanks to Mrs. Ok-Kyung Lim for her technical assistance.

Author information

Authors and Affiliations

  1. Department of Medical Environmental Biology, College of Medicine, Chung-Ang University, Dongjak-gu, Seoul, 156-756, Republic of Korea

    Shunyu Li, Jung Guk Shin, Tae Im Kim & Sung-Jong Hong

  2. Department of Infection Biology, College of Medicine, Wonkwang University, Iksan, Jeonbuk, 570-749, Republic of Korea

    Pyo Yun Cho

  3. Department of Parasitology and Tropical Medicine, College of Medicine, Seoul National University, Jongro-gu, Seoul, 110-799, Republic of Korea

    Sung-Tae Hong

Authors
  1. Shunyu Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jung Guk Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pyo Yun Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tae Im Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sung-Tae Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sung-Jong Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sung-Jong Hong.

Additional information

The nucleotide sequence reported herein was submitted to GenBank and assigned Accession number AY519356.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, S., Shin, J.G., Cho, P.Y. et al. Multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis. Parasitol Res 108, 1295–1302 (2011). https://doi.org/10.1007/s00436-010-2179-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-010-2179-1

Keywords

Search

Navigation