- INTRODUCTION
- METHODS
- Serum samples.
- Pre-adsorption of serum samples for immunoscreening.
- Indirect ELISA.
- cDNA library immunoscreening.
- In vivo excision, plasmid extraction, and sequencing analysis.
- Expression and purification of recombinant protein.
- Protein verification by matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF).
- Evaluation of the antigenicity of the recombinant protein by IgG4-ELISA.
- RESULTS
- DISCUSSION
- CONCLUSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
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: 138–140.
-
3.
Keiser PB & Nutman TB 2004. Strongyloides stercoralis in the immunocompromised population. Clin Microbiol Rev 17: 208–217.
-
4.
Simon GL 2009. Strongyloidiasis. Satoskar AR, Simon GL, Hotez PJ, Tsuji M, eds. Medical Parasitology. Austin, TX: Landes Bioscience, 31–38.
-
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: 967–972.
-
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.
-
7.
Prendki V, Fenaux P, Durand R, Thellier M, Bouchaud O, 2011. Strongyloidiasis in man 75 years after initial exposure. Emerg Infect Dis 17: 931–932.
-
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: 163–167.
-
9.
Iriemenam NC, Sanyaolu AO, Oyibo WA & Fagbenro-Beyioku AF 2010. Strongyloides stercoralis and the immune response. Parasitol Int 59: 9–14.
-
10.
Ramachandran S, Thompson RW, Gam AA & Neva FA 1998. Recombinant cDNA clones for immunodiagnosis of strongyloidiasis. J Infect Dis 177: 196–203.
-
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: 73–81.
-
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: 732–736.
-
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: 174–179.
-
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: 60–66.
-
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: 144–150.
-
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: 272–275.
-
17.
Li J, Zhang WB & McManus DP 2004. Recombinant antigens for immunodiagnosis of cystic echinococcosis. Biol Proced Online 6: 67–77.
-
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: 279–283.
-
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: 24–29.
-
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: 17–26.
-
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: 444–451.
-
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.
-
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: 517–526.
-
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: 34–41.
-
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: 346–350.
-
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: 784–787.
-
27.
Conway DJ, Lindo JF, Robinson RD, Bundy DA & Bianco AE 1994. Strongyloides stercoralis: characterization of immunodiagnostic larval antigens. Exp Parasitol 79: 99–105.
-
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: 609–613.
-
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: 1209–1214.
-
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: 668–671.
-
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: 780–788.
-
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: 4327–4334.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 537 | 323 | 21 |
| Full Text Views | 820 | 7 | 0 |
| PDF Downloads | 220 | 11 | 0 |
Identification and Preliminary Evaluation of a Novel Recombinant Protein for Serodiagnosis of Strongyloidiasis
Norsyahida Arifin
Norsyahida Arifin
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia;
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Muhammad Hafiznur Yunus
Muhammad Hafiznur Yunus
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia;
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Thomas J. Nolan
Thomas J. Nolan
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania
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James B. Lok
James B. Lok
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania
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Rahmah Noordin
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
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.
- INTRODUCTION
- METHODS
- Serum samples.
- Pre-adsorption of serum samples for immunoscreening.
- Indirect ELISA.
- cDNA library immunoscreening.
- In vivo excision, plasmid extraction, and sequencing analysis.
- Expression and purification of recombinant protein.
- Protein verification by matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF).
- Evaluation of the antigenicity of the recombinant protein by IgG4-ELISA.
- RESULTS
- DISCUSSION
- CONCLUSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
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: 138–140.
-
3.
Keiser PB & Nutman TB 2004. Strongyloides stercoralis in the immunocompromised population. Clin Microbiol Rev 17: 208–217.
-
4.
Simon GL 2009. Strongyloidiasis. Satoskar AR, Simon GL, Hotez PJ, Tsuji M, eds. Medical Parasitology. Austin, TX: Landes Bioscience, 31–38.
-
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: 967–972.
-
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.
-
7.
Prendki V, Fenaux P, Durand R, Thellier M, Bouchaud O, 2011. Strongyloidiasis in man 75 years after initial exposure. Emerg Infect Dis 17: 931–932.
-
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: 163–167.
-
9.
Iriemenam NC, Sanyaolu AO, Oyibo WA & Fagbenro-Beyioku AF 2010. Strongyloides stercoralis and the immune response. Parasitol Int 59: 9–14.
-
10.
Ramachandran S, Thompson RW, Gam AA & Neva FA 1998. Recombinant cDNA clones for immunodiagnosis of strongyloidiasis. J Infect Dis 177: 196–203.
-
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: 73–81.
-
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: 732–736.
-
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: 174–179.
-
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: 60–66.
-
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: 144–150.
-
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: 272–275.
-
17.
Li J, Zhang WB & McManus DP 2004. Recombinant antigens for immunodiagnosis of cystic echinococcosis. Biol Proced Online 6: 67–77.
-
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: 279–283.
-
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: 24–29.
-
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: 17–26.
-
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: 444–451.
-
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.
-
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: 517–526.
-
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: 34–41.
-
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: 346–350.
-
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: 784–787.
-
27.
Conway DJ, Lindo JF, Robinson RD, Bundy DA & Bianco AE 1994. Strongyloides stercoralis: characterization of immunodiagnostic larval antigens. Exp Parasitol 79: 99–105.
-
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: 609–613.
-
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: 1209–1214.
-
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: 668–671.
-
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: 780–788.
-
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: 4327–4334.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 537 | 323 | 21 |
| Full Text Views | 820 | 7 | 0 |
| PDF Downloads | 220 | 11 | 0 |