Authors:
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- Introduction
- Materials and Methods
- Collection of T. cati second-stage larvae.
- Human serum samples.
- RNA extraction.
- Complementary DNA synthesis and polymerase chain reaction amplification.
- Cloning of the T. cati TES-120 gene.
- Sequence analysis.
- Subcloning of the T. cati TES-120 gene into an expression vector.
- Expression and purification of T. cati TES-120 recombinant antigen.
- Determination of the antigenicity of the recombinant antigen by western blotting.
- Determination of diagnostic sensitivity and specificity.
- Results
- Discussion
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Magnaval JF, Glickman LT, Dorchies P, Morassin B, 2001. Highlights of human toxocariasis. Korean J Parasitol 39: 1–11.
-
2.
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.
-
3.
Fisher M, 2003. Toxocara cati: an underestimated zoonotic agent. Trends Parasitol 19: 167–170.
-
4.
Smith H, Noordin R, 2006. Diagnostic limitations and future trends in the serodiagnosis of human toxocariasis. Holland CV, Smith HV, eds. Toxocara: The Enigmatic Parasite. Oxfordshire, United Kingdom: CABI Publishing, 89–112.
-
5.
Overgaauw PAM, Nederland V, 1997. Aspects of Toxocara epidemiology: human toxocarosis. Crit Rev Microbiol 23: 215–231.
-
6.
Jansen J, Knapen FV, 1993. Toxocara eggs in public parks and sand-boxes in Utrecht. Tijdschr Diergeneeskd 118: 611–614.
-
7.
Overgaauw PAM, Knapen FV, 2008. Toxocarosis, an important zoonosis. European J Companion Animal Practice 18: 259–266.
-
8.
Wilkins PP, 2014. Immunodiagnosis of human toxocariasis and prospects for improved diagnostics. Curr Trop Med Rep 1: 44–51.
-
9.
Xinou E, Lefkopoulos A, Gelagoti M, Drevelegas A, Diakou A, Milonas I, Dimitriadis AS, 2003. CT and MR imaging findings in cerebral toxocaral disease. AJNR Am J Neuroradiol 24: 714–718.
-
10.
Lee AC, Schantz PM, Kazacos KR, Montgomery SP, Bowman DD, 2010. Epidemiologic and zoonotic aspects of ascarid infections in dogs and cats. Trends Parasitol 26: 155–161.
-
11.
Yamasaki H, Araki K, Lim PK, Zasmy N, Mak JW, Taib R, Aoki T, 2000. Development of a highly specific recombinant Toxocara canis second-stage larva excretory-secretory antigen for immunodiagnosis of human toxocariasis. J Clin Microbiol 38: 1409–1413.
-
12.
Zibaei M, Sadjjadi SM, Sarkari B, 2007. Prevalence of Toxocara cati and other intestinal helminths in stray cats in Shiraz, Iran. Trop Biomed 24: 39–43.
-
13.
Alcântara-Neves NM, dos Santos AB, Mendonça LR, Figueiredo CA, Pontes-de-Carvalho L, 2008. An improved method to obtain antigen-excreting Toxocara canis larvae. Exp Parasitol 119: 349–351.
-
14.
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.
-
15.
Zibaei M, Sadjjadi SM, Jahadi-Hosseini SH, 2014. Toxocara cati larvae in the eye of a child: a case report. Asian Pac J Trop Biomed 4: S53–S55.
-
16.
Petithory JC, Chaumeil C, Liotet S, Rosseau M, Bisognani C, 1993. Immunological studies on ocular larva migrans. Lewis JW, Maizels RM, eds. Toxocara and Toxocariasis: Clinical, Epidemiological and Molecular Perspectives. London, United Kingdom: The British Society for Parasitology with the Institute of Biology, 181–189.
-
17.
Sakai R, Kawashima H, Shibui H, Kamata K, Kambara C, Matsuoka H, 1998. Toxocara cati-induced ocular toxocariasis. Arch Ophthalmol 116: 1686–1687.
-
18.
Shimokawa H, 1982. Visceral larva migrans by Toxocara cati. Fukuoka Igaku Zasshi 73: 64–69.
-
19.
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.
-
20.
Maizels RM, 2013. Toxocara canis: molecular basis of immune recognition and evasion. Vet Parasitol 193: 365–374.
-
21.
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.
-
22.
Kennedy MW, Maizels RM, Meghji M, Young L, Qureshi F, Smith HV, 1987. Species-specific and common epitopes on the secreted and surface antigens of Toxocara cati and Toxocara canis infective larvae. Parasite Immunol 9: 407–420.
-
23.
Poulsen CS, Skov A, Yoshida A, Skallerup P, Maruyama H, Thamsborg SM, Nejsum P, 2015. Differential serodiagnostics of Toxocara canis and T. cati—is it possible? Parasite Immunol. doi:10.1111/pim.12181.
-
24.
Speiser F, Gottstein B, 1984. A collaborative study on larval excretory/secretory antigens of Toxocara canis for the immunodiagnosis of human toxocariasis with ELISA. Acta Trop 41: 361–372.
-
25.
Fong MY, Lau YL, Init I, Jamaiah I, Anuar AK, Noordin R, 2003. Recombinant expression of Toxocara canis excretory-secretory antigen TES-120 in Escherichia coli. Southeast Asian J Trop Med Public Health 34: 723–726.
-
26.
Fong MY, Lau YL, 2004. Recombinant expression of the larval excretory-secretory antigen TES-120 of Toxocara canis in the methylotrophic yeast Pichia pastoris. Parasitol Res 92: 173–176.
-
27.
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.
Production of Toxocara cati TES-120 Recombinant Antigen and Comparison with its T. canis Homolog for Serodiagnosis of Toxocariasis
Farzaneh Zahabiun
Farzaneh Zahabiun
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
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Seyed Mahmoud Sadjjadi
Seyed Mahmoud Sadjjadi
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
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Muhammad Hafiznur Yunus
Muhammad Hafiznur Yunus
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
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Anizah Rahumatullah
Anizah Rahumatullah
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
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Mohammad Hosein Falaki Moghaddam
Mohammad Hosein Falaki Moghaddam
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
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Syazwan Saidin
Syazwan Saidin
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
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Rahmah Noordin
Rahmah Noordin
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran
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Toxocariasis is a cosmopolitan zoonotic disease caused by the infective larvae of Toxocara canis and T. cati. Diagnosis in humans is usually based on clinical symptoms and serology. Immunoglobulin G (IgG)-enzyme-linked immunosorbent assay kits using T. canis excretory–secretory (TES) larval antigens are commonly used for serodiagnosis. Differences in the antigens of the two Toxocara species may influence the diagnostic sensitivity of the test. In this study, T. cati recombinant TES-120 (rTES-120) was cloned, expressed, and compared with its T. canis homolog in an IgG4-western blot. The diagnostic sensitivity and specificity of T. cati rTES-120 were 70% (33/47) and 100% (39/39), respectively. T. canis rTES-120 showed 57.4% sensitivity and 94.4% specificity. When the results of assays using rTES-120 of both species were considered, the diagnostic sensitivity was 76%. This study shows that using antigens from both Toxocara species may improve the serodiagnosis of toxocariasis.
Author Notes
Financial support: This study was funded by the following research grants: Malaysian Cradle Fund no. 304.CIPPM.650564.C117, USM Short Term Grant no. 304.CIPPM.6311089, and SUMS Grant no. 5790.
Authors' addresses: Farzaneh Zahabiun and Seyed Mahmoud Sadjjadi, Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran, E-mails: zahabiun@gmail.com and smsadjjadi@sums.ac.ir. Muhammad Hafiznur Yunus, Anizah Rahumatullah, Syazwan Saidin, and Rahmah Noordin, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia, E-mails: mhn1016@yahoo.com, anirah82@gmail.com, syazwan_saidin@yahoo.com, and rahmah8485@gmail.com. Falaki Moghaddam, MH Department of Medical Parasitology and Mycology, School of Public Health and National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran, E-mail: falakimoghaddam@yahoo.com.
Reprint requests: Rahmah Noordin, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Penang, Malaysia, E-mail: rahmah8485@gmail.com, Tel: +6046534854, or Seyed Mahmoud Sadjjadi, Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran, E-mail: smsadjjadi@sums.ac.ir.
- Introduction
- Materials and Methods
- Collection of T. cati second-stage larvae.
- Human serum samples.
- RNA extraction.
- Complementary DNA synthesis and polymerase chain reaction amplification.
- Cloning of the T. cati TES-120 gene.
- Sequence analysis.
- Subcloning of the T. cati TES-120 gene into an expression vector.
- Expression and purification of T. cati TES-120 recombinant antigen.
- Determination of the antigenicity of the recombinant antigen by western blotting.
- Determination of diagnostic sensitivity and specificity.
- Results
- Discussion
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Magnaval JF, Glickman LT, Dorchies P, Morassin B, 2001. Highlights of human toxocariasis. Korean J Parasitol 39: 1–11.
-
2.
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.
-
3.
Fisher M, 2003. Toxocara cati: an underestimated zoonotic agent. Trends Parasitol 19: 167–170.
-
4.
Smith H, Noordin R, 2006. Diagnostic limitations and future trends in the serodiagnosis of human toxocariasis. Holland CV, Smith HV, eds. Toxocara: The Enigmatic Parasite. Oxfordshire, United Kingdom: CABI Publishing, 89–112.
-
5.
Overgaauw PAM, Nederland V, 1997. Aspects of Toxocara epidemiology: human toxocarosis. Crit Rev Microbiol 23: 215–231.
-
6.
Jansen J, Knapen FV, 1993. Toxocara eggs in public parks and sand-boxes in Utrecht. Tijdschr Diergeneeskd 118: 611–614.
-
7.
Overgaauw PAM, Knapen FV, 2008. Toxocarosis, an important zoonosis. European J Companion Animal Practice 18: 259–266.
-
8.
Wilkins PP, 2014. Immunodiagnosis of human toxocariasis and prospects for improved diagnostics. Curr Trop Med Rep 1: 44–51.
-
9.
Xinou E, Lefkopoulos A, Gelagoti M, Drevelegas A, Diakou A, Milonas I, Dimitriadis AS, 2003. CT and MR imaging findings in cerebral toxocaral disease. AJNR Am J Neuroradiol 24: 714–718.
-
10.
Lee AC, Schantz PM, Kazacos KR, Montgomery SP, Bowman DD, 2010. Epidemiologic and zoonotic aspects of ascarid infections in dogs and cats. Trends Parasitol 26: 155–161.
-
11.
Yamasaki H, Araki K, Lim PK, Zasmy N, Mak JW, Taib R, Aoki T, 2000. Development of a highly specific recombinant Toxocara canis second-stage larva excretory-secretory antigen for immunodiagnosis of human toxocariasis. J Clin Microbiol 38: 1409–1413.
-
12.
Zibaei M, Sadjjadi SM, Sarkari B, 2007. Prevalence of Toxocara cati and other intestinal helminths in stray cats in Shiraz, Iran. Trop Biomed 24: 39–43.
-
13.
Alcântara-Neves NM, dos Santos AB, Mendonça LR, Figueiredo CA, Pontes-de-Carvalho L, 2008. An improved method to obtain antigen-excreting Toxocara canis larvae. Exp Parasitol 119: 349–351.
-
14.
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.
-
15.
Zibaei M, Sadjjadi SM, Jahadi-Hosseini SH, 2014. Toxocara cati larvae in the eye of a child: a case report. Asian Pac J Trop Biomed 4: S53–S55.
-
16.
Petithory JC, Chaumeil C, Liotet S, Rosseau M, Bisognani C, 1993. Immunological studies on ocular larva migrans. Lewis JW, Maizels RM, eds. Toxocara and Toxocariasis: Clinical, Epidemiological and Molecular Perspectives. London, United Kingdom: The British Society for Parasitology with the Institute of Biology, 181–189.
-
17.
Sakai R, Kawashima H, Shibui H, Kamata K, Kambara C, Matsuoka H, 1998. Toxocara cati-induced ocular toxocariasis. Arch Ophthalmol 116: 1686–1687.
-
18.
Shimokawa H, 1982. Visceral larva migrans by Toxocara cati. Fukuoka Igaku Zasshi 73: 64–69.
-
19.
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.
-
20.
Maizels RM, 2013. Toxocara canis: molecular basis of immune recognition and evasion. Vet Parasitol 193: 365–374.
-
21.
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.
-
22.
Kennedy MW, Maizels RM, Meghji M, Young L, Qureshi F, Smith HV, 1987. Species-specific and common epitopes on the secreted and surface antigens of Toxocara cati and Toxocara canis infective larvae. Parasite Immunol 9: 407–420.
-
23.
Poulsen CS, Skov A, Yoshida A, Skallerup P, Maruyama H, Thamsborg SM, Nejsum P, 2015. Differential serodiagnostics of Toxocara canis and T. cati—is it possible? Parasite Immunol. doi:10.1111/pim.12181.
-
24.
Speiser F, Gottstein B, 1984. A collaborative study on larval excretory/secretory antigens of Toxocara canis for the immunodiagnosis of human toxocariasis with ELISA. Acta Trop 41: 361–372.
-
25.
Fong MY, Lau YL, Init I, Jamaiah I, Anuar AK, Noordin R, 2003. Recombinant expression of Toxocara canis excretory-secretory antigen TES-120 in Escherichia coli. Southeast Asian J Trop Med Public Health 34: 723–726.
-
26.
Fong MY, Lau YL, 2004. Recombinant expression of the larval excretory-secretory antigen TES-120 of Toxocara canis in the methylotrophic yeast Pichia pastoris. Parasitol Res 92: 173–176.
-
27.
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.