- INTRODUCTION
- MATERIALS AND METHODS
- Serum samples.
- Testing serum samples with commercial IgG-ELISA kits.
- Recombinant protein expression and purification.
- MALDI TOF/TOF analysis of the recombinant protein.
- Western blotting recombinant proteins.
- Development of the lateral flow dipstick test.
- Preparation of the lateral flow dipstick.
- Optimization of the antigen concentration and the OD for the anti-human IgG4 colloidal gold conjugate.
- Lateral flow dipstick test procedure.
- RESULTS
- DISCUSSION
- CONCLUSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Pawlowski Z, 2001. Toxocariasis in humans: clinical expression and treatment dilemma. J Helminthol 75: 299–305.
-
2.
Glickman LT, Schantz PM, 1981. Epidemiology and pathogenesis of zoonotic toxocariasis. Epidemiol Rev 3: 230–250.
-
3.
Fialho PM, Correa CR, 2016. A systematic review of toxocariasis: a neglected but high-prevalence disease in Brazil. Am J Trop Med Hyg 94: 1193–1199.
-
4.
Smith H, Noordin R, 2006. Diagnostic limitations and future trends in the serodiagnosis of human toxocariasis. Holland C, Smith HV, eds. Toxocara: The Enigmatic Parasite. Wallingford, United Kingdom: CABI Publishing, 93–102.
-
5.
Hotez PJ, Wilkins PP, 2009. Toxocariasis: America’s most common neglected infection of poverty and a helminthiasis of global importance? PLoS Negl Trop Dis 3: e400.
-
6.
Mohamad S, Azmi NC, Noordin R, 2009. Development and evaluation of a sensitive and specific assay for diagnosis of human toxocariasis by use of three recombinant antigens (TES-26, TES-30USM, and TES-120). J Clin Microbiol 47: 1712–1717.
-
7.
Yamasaki H, Taib R, Watanabe Y-i, Mak JW, Zasmy N, Araki K, Chooi LPK, Kita K, Aoki T, 1998. Molecular characterization of a cDNA encoding an excretory–secretory antigen from Toxocara canis second stage larvae and its application to the immunodiagnosis of human toxocariasis1. Parasitol Int 47: 171–181.
-
8.
Smith H, Holland C, Taylor M, Magnaval JF, Schantz P, Maizels R, 2009. How common is human toxocariasis? Towards standardizing our knowledge. Trends Parasitol 25: 182–188.
-
9.
Cypess RH, Karol MH, Zidian JL, Glickman LT, Gitlin D, 1977. Larva-specific antibodies in patients with visceral larva migrans. J Infect Dis 135: 633–640.
-
10.
Roldan WH, Espinoza YA, 2009. Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis. Mem Inst Oswaldo Cruz 104: 411–418.
-
11.
Sambrook J, Russell DW, 2001. Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press.
-
12.
Saidin S, Yunus MH, Zakaria ND, Razak KA, Huat LB, Othman N, Noordin R, 2014. Production of recombinant Entamoeba histolytica pyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess. BMC Infect Dis 14: 182.
-
13.
Perkins DN, Pappin DJ, Creasy DM, Cottrell JS, 1999. Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20: 3551–3567.
-
14.
Makhsin SR, Razak KA, Noordin R, Zakaria ND, Chun TS, 2012. The effects of size and synthesis methods of gold nanoparticle-conjugated MalphaHIgG4 for use in an immunochromatographic strip test to detect brugian filariasis. Nanotechnology 23: 495719.
-
15.
Jacquier P, Gottstein B, Stingelin Y, Eckert J, 1991. Immunodiagnosis of toxocarosis in humans: evaluation of a new enzyme-linked immunosorbent assay kit. J Clin Microbiol 29: 1831–1835.
-
16.
Magnaval JF, Glickman LT, Dorchies P, Morassin B, 2001. Highlights of human toxocariasis. Korean J Parasitol 39: 1–11.
-
17.
Camargo ED, Nakamura PM, Vaz AJ, da Silva MV, Chieffi PP, de Melo EO, 1992. Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparison of the assay with ELISA. Rev Inst Med Trop São Paulo 34: 55–60.
-
18.
Ishida MM, Rubinsky-Elefant G, Ferreira AW, Hoshino-Shimizu S, Vaz AJ, 2003. Helminth antigens (Taenia solium, Taenia crassiceps, Toxocara canis, Schistosoma mansoni and Echinococcus granulosus) and cross-reactivities in human infections and immunized animals. Acta Trop 89: 73–84.
-
19.
Magnaval JF, Fabre R, Maurieres P, Charlet JP, de Larrard B, 1991. Application of the western blotting procedure for the immunodiagnosis of human toxocariasis. Parasitol Res 77: 697–702.
-
20.
Noordin R, Smith HV, Mohamad S, Maizels RM, Fong MY, 2005. Comparison of IgG-ELISA and IgG4-ELISA for Toxocara serodiagnosis. Acta Trop 93: 57–62.
-
21.
Fillaux J, Magnaval JF, 2013. Laboratory diagnosis of human toxocariasis. Vet Parasitol 193: 327–336.
-
22.
Zhan B et al. 2015. Identification of immunodominant antigens for the laboratory diagnosis of toxocariasis. Trop Med Int Health 20: 1787–1796.
-
23.
Peeling RW, Holmes KK, Mabey D, Ronald A, 2006. Rapid tests for sexually transmitted infections (STIs): the way forward. Sex Transm Infect 82 (Suppl 5): v1–v6.
-
24.
Akao N, Chu AE, Tsukidate S, Fujita K, 1997. A rapid and sensitive screening kit for the detection of anti-Toxocara larval ES antibodies. Parasitol Int 46: 189–195.
-
25.
Bojanich MV, Marino GL, López MÁ, Alonso JM, 2012. An evaluation of the dot-ELISA procedure as a diagnostic test in an area with a high prevalence of human Toxocara canis infection. Mem Inst Oswaldo Cruz 107: 194–197.
-
26.
Lim PK, Yamasaki H, Mak JW, Wong SF, Chong CW, Yap IK, Ambu S, Kumarasamy V, 2015. Field evaluation of a rapid diagnostic test to detect antibodies in human toxocariasis. Acta Trop 148: 32–37.
-
27.
Kwan-Lim GE, Forsyth KP, Maizels RM, 1990. Filarial-specific IgG4 response correlates with active Wuchereria bancrofti infection. J Immunol 145: 4298–4305.
-
28.
Lucius R, Kern A, Seeber F, Pogonka T, Willenbucher J, Taylor HR, Pinder M, Ghalib HW, Schulz-Key H, Soboslay P, 1992. Specific and sensitive IgG4 immunodiagnosis of onchocerciasis with a recombinant 33 kD Onchocerca volvulus protein (Ov33). Trop Med Parasitol 43: 139–145.
-
29.
Egwang TG, Duong TH, Nguiri C, Ngari P, Everaere S, Richard-Lenoble D, Gbakima AA, Kombila M, 1994. Evaluation of Onchocerca volvulus-specific IgG4 subclass serology as an index of onchocerciasis transmission potential of three Gabonese villages. Clin Exp Immunol 98: 401–407.
-
31.
Weil GJ, Ogunrinade AF, Chandrashekar R, Kale OO, 1990. IgG4 subclass antibody serology for onchocerciasis. J Infect Dis 161: 549–554.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 307 | 244 | 38 |
| Full Text Views | 570 | 9 | 1 |
| PDF Downloads | 143 | 7 | 1 |
A Lateral Flow Rapid Test for Human Toxocariasis Developed Using Three Toxocara canis Recombinant Antigens
Muhammad Hafiznur Yunus
Muhammad Hafiznur Yunus
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia
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Siti Naqiuyah Tan Farrizam
Siti Naqiuyah Tan Farrizam
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia
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Izzati Zahidah Abdul Karim
Izzati Zahidah Abdul Karim
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia
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Rahmah Noordin
Rahmah Noordin
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia
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Laboratory diagnosis of toxocariasis is still a challenge especially in developing endemic countries with polyparasitism. In this study, three Toxocara canis recombinant antigens, rTES-26, rTES-30, and rTES-120, were expressed and used to prepare lateral flow immunoglobulin G4 (IgG4) dipsticks. The concordance of the results of the rapid test (comprising three dipsticks) with a commercial IgG-enzyme-linked immunosorbent assay (ELISA) (Cypress Diagnostics, Belgium) was compared against the concordance of two other commercial IgG-ELISA kits (Bordier, Switzerland and NovaTec, Germany) with the Cypress kit. Using Toxocara-positive samples, the concordance of the dipstick dotted with rTES-26, rTES-30, and rTES-120 was 41.4% (12/29), 51.7% (15/29), and 72.4% (21/29), respectively. When positivity with any dipstick was considered as an overall positive rapid test result, the concordance with the Cypress kit was 93% (27/29). Meanwhile, when compared with the results of the Cypress kit, the concordance of IgG-ELISA from NovaTec and Bordier was 100% (29/29) and 89.7% (26/29), respectively. Specific IgG4 has been recognized as a marker of active infection for several helminthic diseases; therefore, the two non-concordant results of the rapid test when compared with the NovaTec IgG-ELISA kit may be from samples of people with non-active infection. All the three dipsticks showed 100% (50/50) concordance with the Cypress kit when tested with serum from individuals who were healthy and with other infections. In conclusion, the lateral flow rapid test is potentially a good, fast, and easy test for toxocariasis. Next, further validation studies and development of a test with the three antigens in one dipstick will be performed.
Author Notes
Financial support: The study was funded by Universiti Sains Malaysia Short Term grant (Grant no: 304.CIPPM.6311089), Cradle Fund Sdn. Bhd. (Grant no: 304.CIPPM.650564.C117), and Universiti Sains Malaysia Research University grant (Grant no: 1001.CIPPM.8012202).
Authors’ addresses: Muhammad Hafiznur Yunus, Siti Naqiuyah Tan Farrizam, Izzati Zahidah Abdul Karim, and Rahmah Noordin, Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang, Malaysia, E-mails: mhn1016@yahoo.com, naddytan.f@gmail.com, ezzati87@gmail.com, and rahmah8485@gmail.com.
- INTRODUCTION
- MATERIALS AND METHODS
- Serum samples.
- Testing serum samples with commercial IgG-ELISA kits.
- Recombinant protein expression and purification.
- MALDI TOF/TOF analysis of the recombinant protein.
- Western blotting recombinant proteins.
- Development of the lateral flow dipstick test.
- Preparation of the lateral flow dipstick.
- Optimization of the antigen concentration and the OD for the anti-human IgG4 colloidal gold conjugate.
- Lateral flow dipstick test procedure.
- RESULTS
- DISCUSSION
- CONCLUSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Pawlowski Z, 2001. Toxocariasis in humans: clinical expression and treatment dilemma. J Helminthol 75: 299–305.
-
2.
Glickman LT, Schantz PM, 1981. Epidemiology and pathogenesis of zoonotic toxocariasis. Epidemiol Rev 3: 230–250.
-
3.
Fialho PM, Correa CR, 2016. A systematic review of toxocariasis: a neglected but high-prevalence disease in Brazil. Am J Trop Med Hyg 94: 1193–1199.
-
4.
Smith H, Noordin R, 2006. Diagnostic limitations and future trends in the serodiagnosis of human toxocariasis. Holland C, Smith HV, eds. Toxocara: The Enigmatic Parasite. Wallingford, United Kingdom: CABI Publishing, 93–102.
-
5.
Hotez PJ, Wilkins PP, 2009. Toxocariasis: America’s most common neglected infection of poverty and a helminthiasis of global importance? PLoS Negl Trop Dis 3: e400.
-
6.
Mohamad S, Azmi NC, Noordin R, 2009. Development and evaluation of a sensitive and specific assay for diagnosis of human toxocariasis by use of three recombinant antigens (TES-26, TES-30USM, and TES-120). J Clin Microbiol 47: 1712–1717.
-
7.
Yamasaki H, Taib R, Watanabe Y-i, Mak JW, Zasmy N, Araki K, Chooi LPK, Kita K, Aoki T, 1998. Molecular characterization of a cDNA encoding an excretory–secretory antigen from Toxocara canis second stage larvae and its application to the immunodiagnosis of human toxocariasis1. Parasitol Int 47: 171–181.
-
8.
Smith H, Holland C, Taylor M, Magnaval JF, Schantz P, Maizels R, 2009. How common is human toxocariasis? Towards standardizing our knowledge. Trends Parasitol 25: 182–188.
-
9.
Cypess RH, Karol MH, Zidian JL, Glickman LT, Gitlin D, 1977. Larva-specific antibodies in patients with visceral larva migrans. J Infect Dis 135: 633–640.
-
10.
Roldan WH, Espinoza YA, 2009. Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis. Mem Inst Oswaldo Cruz 104: 411–418.
-
11.
Sambrook J, Russell DW, 2001. Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press.
-
12.
Saidin S, Yunus MH, Zakaria ND, Razak KA, Huat LB, Othman N, Noordin R, 2014. Production of recombinant Entamoeba histolytica pyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess. BMC Infect Dis 14: 182.
-
13.
Perkins DN, Pappin DJ, Creasy DM, Cottrell JS, 1999. Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20: 3551–3567.
-
14.
Makhsin SR, Razak KA, Noordin R, Zakaria ND, Chun TS, 2012. The effects of size and synthesis methods of gold nanoparticle-conjugated MalphaHIgG4 for use in an immunochromatographic strip test to detect brugian filariasis. Nanotechnology 23: 495719.
-
15.
Jacquier P, Gottstein B, Stingelin Y, Eckert J, 1991. Immunodiagnosis of toxocarosis in humans: evaluation of a new enzyme-linked immunosorbent assay kit. J Clin Microbiol 29: 1831–1835.
-
16.
Magnaval JF, Glickman LT, Dorchies P, Morassin B, 2001. Highlights of human toxocariasis. Korean J Parasitol 39: 1–11.
-
17.
Camargo ED, Nakamura PM, Vaz AJ, da Silva MV, Chieffi PP, de Melo EO, 1992. Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparison of the assay with ELISA. Rev Inst Med Trop São Paulo 34: 55–60.
-
18.
Ishida MM, Rubinsky-Elefant G, Ferreira AW, Hoshino-Shimizu S, Vaz AJ, 2003. Helminth antigens (Taenia solium, Taenia crassiceps, Toxocara canis, Schistosoma mansoni and Echinococcus granulosus) and cross-reactivities in human infections and immunized animals. Acta Trop 89: 73–84.
-
19.
Magnaval JF, Fabre R, Maurieres P, Charlet JP, de Larrard B, 1991. Application of the western blotting procedure for the immunodiagnosis of human toxocariasis. Parasitol Res 77: 697–702.
-
20.
Noordin R, Smith HV, Mohamad S, Maizels RM, Fong MY, 2005. Comparison of IgG-ELISA and IgG4-ELISA for Toxocara serodiagnosis. Acta Trop 93: 57–62.
-
21.
Fillaux J, Magnaval JF, 2013. Laboratory diagnosis of human toxocariasis. Vet Parasitol 193: 327–336.
-
22.
Zhan B et al. 2015. Identification of immunodominant antigens for the laboratory diagnosis of toxocariasis. Trop Med Int Health 20: 1787–1796.
-
23.
Peeling RW, Holmes KK, Mabey D, Ronald A, 2006. Rapid tests for sexually transmitted infections (STIs): the way forward. Sex Transm Infect 82 (Suppl 5): v1–v6.
-
24.
Akao N, Chu AE, Tsukidate S, Fujita K, 1997. A rapid and sensitive screening kit for the detection of anti-Toxocara larval ES antibodies. Parasitol Int 46: 189–195.
-
25.
Bojanich MV, Marino GL, López MÁ, Alonso JM, 2012. An evaluation of the dot-ELISA procedure as a diagnostic test in an area with a high prevalence of human Toxocara canis infection. Mem Inst Oswaldo Cruz 107: 194–197.
-
26.
Lim PK, Yamasaki H, Mak JW, Wong SF, Chong CW, Yap IK, Ambu S, Kumarasamy V, 2015. Field evaluation of a rapid diagnostic test to detect antibodies in human toxocariasis. Acta Trop 148: 32–37.
-
27.
Kwan-Lim GE, Forsyth KP, Maizels RM, 1990. Filarial-specific IgG4 response correlates with active Wuchereria bancrofti infection. J Immunol 145: 4298–4305.
-
28.
Lucius R, Kern A, Seeber F, Pogonka T, Willenbucher J, Taylor HR, Pinder M, Ghalib HW, Schulz-Key H, Soboslay P, 1992. Specific and sensitive IgG4 immunodiagnosis of onchocerciasis with a recombinant 33 kD Onchocerca volvulus protein (Ov33). Trop Med Parasitol 43: 139–145.
-
29.
Egwang TG, Duong TH, Nguiri C, Ngari P, Everaere S, Richard-Lenoble D, Gbakima AA, Kombila M, 1994. Evaluation of Onchocerca volvulus-specific IgG4 subclass serology as an index of onchocerciasis transmission potential of three Gabonese villages. Clin Exp Immunol 98: 401–407.
-
31.
Weil GJ, Ogunrinade AF, Chandrashekar R, Kale OO, 1990. IgG4 subclass antibody serology for onchocerciasis. J Infect Dis 161: 549–554.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 307 | 244 | 38 |
| Full Text Views | 570 | 9 | 1 |
| PDF Downloads | 143 | 7 | 1 |