Advertisement

Molecular Diagnosis & Therapy

, Volume 22, Issue 4, pp 485–491 | Cite as

Molecular and Immnune Diagnosis: Further Testing for Human Strongyloidiasis

  • Larissa R. Bosqui
  • Priscilla D. Marques
  • Gessica B. de Melo
  • Maria do Rosário F. Gonçalves-Pires
  • Fernanda M. Malta
  • Wander R. Pavanelli
  • Ivete Conchon-Costa
  • Julia M. Costa-Cruz
  • Fabiana M. Paula
  • Idessania N. Costa
Original Research Article
  • 18 Downloads

Abstract

Introduction

Detection of Strongyloides stercoralis larvae is particularly challenging because only a small number of larvae are released into the feces, regardless of infection stage.

Objective

Our objective was to apply conventional polymerase chain reaction (PCR) to the detection of S. stercoralis DNA in feces samples to evaluate its performance in samples of patients with strongyloidiasis and compare results with those of immunodiagnosis.

Methods

Stool, serum, and saliva samples were collected from each individual (n = 48) at the clinic hospital of the State University of Londrina, Brazil, for parasitological, immunological, and molecular tests. Stool samples were processed via parasitological methods. Serum samples were used for immunoglobulin G (IgG) detection and saliva samples for IgA detection by ELISA.

Results

For amplification by conventional PCR, two different primers were used: species specific (101 bp) and genus specific (392 bp). The results showed that 34 (97.1%) of the 35 copro-positive individuals for S. stercoralis were positive for serum IgG and 19 (54.3%) were positive for salivary IgA. Regarding molecular analysis, both primers (species and genus specific) demonstrated positivity in 100% of the samples, which was confirmed by sequencing the positive samples.

Conclusion

Complementary examinations of the parasitological method demonstrated excellent results in the context of the diagnosis of strongyloidiasis, especially in asymptomatic patients with irregular larval release in the feces.

Notes

Compliance with Ethical Standards

Funding

This study was supported by grants from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES—Grant no. 40002012026M9 [LRB] and Fundação de Amparo a Pesquisa do estado de São Paulo (FAPESP—Grant no. 2013/03304-0).

Conflict of interest

LRB, PDM, GBdM, MdRFG-P, FMM, WRP, IC-C, JMC-C, FMP and INC have no conflicts of interest.

Ethical Approval and Informed Consent

The study was approved by the ethics committee on human studies at the State University of Londrina (protocol number 1.306.715).

References

  1. 1.
    Buonfrate D, Sequi M, Mejia R, Cimino RO, Krolewiecki AJ, Albonico M, Degani M, Tais S, Angheben A, Requena-Mendez A, Muñoz J, Nutman TB, Bisoffi Z. Accuracy of five serologic tests for the follow up of Strongyloides stercoralis infection. PLoS Negl Trop Dis. 2015;9:e0003491.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Bisoffi Z, Buonfrate D, Montresor A, Requena-Mendez A, Muñoz J, Krolewiecki AJ, Gotuzzo E, Mena MA, Chiodini PL, Anselmi M, Moreira J, Albonico M. Strongyloides stercoralis: a plea for action. PLoS Negl Trop Dis. 2013;7:e2214.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Olsen A, van Lieshout L, Marti H, Polderman T, Polman K, Steinmann P, Stothard R, Thybo S, Verweij JJ, Magnussen P. Strongyloidiasis—the most neglected of the neglected tropical diseases? Trans R Soc Trop Med Hyg. 2009;103:967–72.CrossRefPubMedGoogle Scholar
  4. 4.
    Nutman TB. Human infection with Strongyloides stercoralis and other related Strongyloides species. Parasitology. 2017;144:263–73.CrossRefPubMedGoogle Scholar
  5. 5.
    Repetto SA, Ruybal P, Solana ME, López C, Berini CA, Alba Soto CD, Cappa SM. Comparison between PCR and larvae visualization methods for diagnosis of Strongyloides stercoralis out of endemic area: a proposed algorithm. Acta Trop. 2016;157:169–77.CrossRefPubMedGoogle Scholar
  6. 6.
    Yap P, Fürst T, Müller I, Kriemler S, Utzinger J, Steinmann P. Determining soil-transmitted helminth infection status and physical fitness of school-aged children. J Vis Exp. 2012;66:e3966.Google Scholar
  7. 7.
    Llewellyn S, Inpankaew T, Nery SV, Gray DJ, Verweij JJ, Clements AC, Gomes SJ, Traub R, McCarthy JS. Application of a multiplex quantitative PCR to assess prevalence and intensity of intestinal parasite infections in a controlled clinical trial. PLoS Negl Trop Dis. 2016;10:e0004380.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Marcos LA, Terashima A, Canales M, Gotuzzo E. Update on strongyloidiasis in the immunocompromised host. Curr Infect Dis Rep. 2011;13:35–46.CrossRefPubMedGoogle Scholar
  9. 9.
    Verweij JJ, Stensvold CR. Molecular testing for clinical diagnosis and epidemiological investigations of intestinal parasitic infections. Clin Microbiol Rev. 2014;27:371–418.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Nilforoushan MR, Mirhendi H, Rezaie S, Rezaian M, Meamar AR, Kia EB. A DNA-based identification of Strongyloides stercoralis isolates from Iran. Iran J Publ Health. 2007;36:16–20.Google Scholar
  11. 11.
    Lodh N, Caro N, Sofer S, Scott A, Krolewiecki A, Shiff C. Diagnóstico de Strongyloides stercoralis: Detecção de DNA derivado de parasita na urina. Acta Trop. 2016;163:9–13.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Dong L, Yoo HB, Wang J, Park SR. Accurate quantification of supercoiled DNA by digital PCR. Sci Rep. 2016;6:24230.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Moghaddassani H, Mirhendi H, Hosseini M, Rokni MB, Mowlavi GH, Kia EB. Molecular diagnosis of Strongyloides stercoralis infection by PCR detection of specific DNA in human stool samples. Iran J Parasitol. 2011;6:23–30.PubMedPubMedCentralGoogle Scholar
  14. 14.
    Paula FM, Malta FM, Marques PD, Sitta RB, Pinho JR, Gryschek RCB, Chieffi PP. Molecular diagnosis of strongyloidiasis in tropical areas: a comparison of conventional and real-time polymerase chain reaction with parasitological methods. Mem. Inst. Oswaldo Cruz 2015;110:272–4.Google Scholar
  15. 15.
    Ahmad AF, Hadip F, Ngui R, Lim YA, Mahmud R. Serological and molecular detection of Strongyloides stercoralis infection among an Orang Asli community in Malaysia. Parasitol Res. 2013;112:2811–6.CrossRefPubMedGoogle Scholar
  16. 16.
    Zueter AM, Mohamed Z, Abdullah AD, Mohamad N, Arifin N, Othman N, Noordi R. Detection of Strongyloides stercoralis infection among cancer patients in a major hospital in Kelantan, Malaysia. Singapore Med J. 2014;55:367–71.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Paula FM, Malta FM, Corral MA, Marques PD, Gottardi M, Meisel DM, Yamashiro J, Pinho JR, Castilho VL, Gonçalves EM, Gryschek RC, Chieffi PP. Diagnosis of Strongyloides stercoralis infection in immunocompromised patients by serological and molecular methods. Rev Inst Med Trop Sao Paulo 2016;58:63.Google Scholar
  18. 18.
    Hoffmann WA, Pons JA, Janer JL. The sedimentation concentration method in schistosomiasis mansoni, Puerto Rico. J Publ Health Trop Med. 1934;9:283–91.Google Scholar
  19. 19.
    Faust EC, Sawitz W, Tobie J, Odon V, Peres C, Lincicome DR. Comparative efficiency of various techniques for the diagnosis of protozoa and helminthes in feces. J Parasitol. 1939;25:241–62.CrossRefGoogle Scholar
  20. 20.
    Kato K, Miura M. Comparative examinations. Jpn J Parasitol. 1954;3:35.Google Scholar
  21. 21.
    Katz N, Chaves A, Pellegrino J. A simple device for quantitative stool thick-smear technique in schistosomiasis mansoni. Rev Inst Med Trop São Paulo. 1972;14:397–400.PubMedGoogle Scholar
  22. 22.
    Rugai E, Mattos T, Brisola AP. Nova técnica para isolar larvas de nematóides das fezes: modificações do método de Baermann. Rev Inst Adolfo Lutz. 1954;14:5–8.PubMedGoogle Scholar
  23. 23.
    Costa IN, Sopelete MC, Gonçalves-Pires MRF, Costa-Cruz JM. IgA and IgG antibodies in paired serum and saliva samples in human strongyloidiasis. Acta Parasitol. 2003;48:306–11.Google Scholar
  24. 24.
    Ribeiro VS, Feliciano ND, Gonzaga HT, Costa IN, Gonçalves-Pires MR, Ueta MT, Costa-Cruz JM. Detergent fraction of heterologous antigen to detect IgA and IgG in strongyloidiasis using saliva and serum paired samples. Immunol Lett. 2010;134:69–74.CrossRefPubMedGoogle Scholar
  25. 25.
    Machado ER, Ueta MT, Gonçalves-Pires MRF, Alves de Oliveira JB, Faccioli LH, Costa-Cruz JM. Strongyloides venezuelensis alkaline extract for the diagnosis of human strongyloidiasis by enzyme-linked immunosorbent assay. Mem Inst Oswaldo Cruz. 2003;98:849–51.CrossRefPubMedGoogle Scholar
  26. 26.
    Verweij JJ, Canales M, Polman K, Ziem J, Brienen EA, Polderman AM, van Lieshout L. Molecular diagnosis of Strongyloides stercoralis in faecal samples using real-time PCR. Trans R Soc Trop Med Hyg. 2009;103:342–6.CrossRefPubMedGoogle Scholar
  27. 27.
    Dorris M, Viney ME, Blaxter LM. Molecular phylogenetic analysis of the genus Strongyloides and related nematodes. Int J Parasitol. 2002;32:1507–17.CrossRefPubMedGoogle Scholar
  28. 28.
    Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci EUA 1977; 74:5463–7.Google Scholar
  29. 29.
    Basuni M, Muhi J, Othman N, Verweij JJ, Ahmad M, Miswan N, Rahumatullah A, Aziz FA, Zainudin NS, Noordin R. A pentaplex real-time polymerase chain reaction assay for detection of four species of soil-transmitted helminths. Am J Trop Med Hyg. 2011;84:338–43.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Taniuchi M, Verweij JJ, Noor Z, Sobuz SU, van Lieshout L, Petri WA Jr, Haque R, Houpt ER. High throughput multiplex PCR and probe-based detection with Luminex beads for seven intestinal parasites. Am J Trop Med Hyg. 2011;84:332–7.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Mendes T, Minori K, Ueta M, Miguel DC, Allegretti SM. Strongyloidiasis current status with emphasis in diagnosis and drug research. J Parasitol Res. 2017;2017:5056314.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Gonçalves ALR, Rocha CA, Gonzaga HT, Gonçalves-Pires MRF, Ueta MT, Costa-Cruz JM. Specific IgG and IgA to larvae, parthenogenetic females and eggs of Strongyloides venezuelensis in the immunodiagnosis of human strongyloidiasis. Diagn Microbiol Infect Dis. 2012;72:79–84.CrossRefPubMedGoogle Scholar
  33. 33.
    Sitta RB, Malta FM, Pinho JR, Chieffi PP, Gryschek RCB, Paula FM. Conventional PCR for molecular diagnosis of human strongyloidiasis. Parasitology. 2014;141:716–21.CrossRefPubMedGoogle Scholar
  34. 34.
    Bosqui LR, Gonçalves AL, Gonçalves-Pires Mdo R, Custodio LA, de Menezes MC, Murad VA, de Paula FM, Pavanelli WR, Conchon-Costa I, Costa-Cruz JM, Costa IN. Detection of parasite-specific IgG and IgA in paired serum and saliva samples for diagnosis of human strongyloidiasis in northern Paraná state, Brazil. Acta Trop. 2015;150:190–5.CrossRefPubMedGoogle Scholar
  35. 35.
    Siddiqui AA, Berk SL. Diagnosis of Strongyloides stercoralis infection. Clin Infect Dis. 2001;33:1040–7.CrossRefPubMedGoogle Scholar
  36. 36.
    Agrawal V, Agarwal T, Ghoshal UC. Intestinal strongyloidiasis: a diagnosis frequently missed in the tropics. Trans R Soc Trop Med Hyg. 2009;103:242–6.CrossRefPubMedGoogle Scholar
  37. 37.
    Uparanukraw P, Phongsri S, Morakote N. Fluctuations of larval excretion in Strongyloides stercoralis infection. Am J Trop Med Hyg. 1999;60:967–73.CrossRefPubMedGoogle Scholar
  38. 38.
    Johnston FH, Morris PS, Speare R, McCarthy J, Currie B, Ewald D, Page W, Dempsey K. Strongyloidiasis: a review of the evidence for Australian practitioners. Aust J Rural Health. 2005;13:247–54.CrossRefPubMedGoogle Scholar
  39. 39.
    Atkins NS, Lindo JF, Lee MG, Conway DJ, Bailey JW, Robinson RD, Bundy DA. Humoral responses in human strongyloidiasis: correlations with infection chronicity. Trans R Soc Trop Med Hyg. 1997;91:609–13.CrossRefPubMedGoogle Scholar
  40. 40.
    Inês EJ, Silva MLS, Souza JN, Teixeira MCA, Soares NM. The role of glycosylated epitopes in the serodiagnosis of Strongyloides stercoralis infection. Diagn Microbiol Infect Dis. 2013;76:31–5.CrossRefGoogle Scholar
  41. 41.
    Khare R, Espy MJ, Cebelinski E, Boxrud D, Sloan LM, Cunningham SA, Pritt BS, Patel R, Binnicker MJ. Comparative evaluation of two commercial multiplex panels for detection of gastrointestinal pathogens by use of clinical stool specimens. J Clin Microbiol. 2014;52:3667–73.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Buonfrate D, Requena-Mendez A, Angheben A, Cinquini M, Cruciani M, Fittipaldo A, Giorli G, Gobbi F, Piubelli C, Bisoffi Z. Accuracy of molecular biology techniques for the diagnosis of Strongyloides stercoralis infection—a systematic review and meta-analysis. PLoS Negl Trop Dis. 2018;12:e0006229.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Boggild AK, Libman M, Greenaway CA, McCarthy AE. CATMAT Statement on disseminated strongyloidiasis: prevention, assessment and management guidelines. Can Commun Dis Rep. 2016;42:12–9.CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Janwan P, Intapan PM, Thanchomnang T, Lulitanond V, Anamnart W, Maleewong W. Rapid detection of Opisthorchis viverrini and Strongyloides stercoralis in human fecal samples using a duplex real-time PCR and melting curve analysis. Parasitol Res. 2011;109:1593–601.CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Larissa R. Bosqui
    • 1
  • Priscilla D. Marques
    • 2
  • Gessica B. de Melo
    • 2
  • Maria do Rosário F. Gonçalves-Pires
    • 3
  • Fernanda M. Malta
    • 2
  • Wander R. Pavanelli
    • 1
  • Ivete Conchon-Costa
    • 1
  • Julia M. Costa-Cruz
    • 3
  • Fabiana M. Paula
    • 2
  • Idessania N. Costa
    • 1
  1. 1.Departamento de Ciências Patológicas, CCB, Laboratório de Parasitologia ExperimentalUniversidade Estadual de LondrinaLondrinaBrazil
  2. 2.Departamento de Moléstias Infecciosas e Parasitárias - Hospital das Clínicas da Faculdade de Medicina da Universidade de São PauloSão PauloBrazil
  3. 3.Departamento de Parasitologia, Instituto de Ciências BiomédicasUniversidade Federal de UberlândiaUberlândiaBrazil

Personalised recommendations