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Approaches to Detecting Cyclospora cayetanensis: From Diagnostics to Detection in Different Food Matrices

  • Ynés R. Ortega
  • Lucy J. Robertson
Chapter
Part of the SpringerBriefs in Food, Health, and Nutrition book series (BRIEFSFOOD)

Abstract

Detection of Cyclospora oocysts in feces is the gold standard of clinical diagnostics, although efficient multiplex diagnostic kits relying on molecular techniques are adding to the armory. However, in order to be able to link cases, confirm transmission vehicles (often implicated epidemiologically, but sometimes with relatively flimsy evidence), and determine routes from source to infection, we need also to be able to analyze environmental matrices, particularly water samples and food, for Cyclospora. Our detection tools, be they based on molecular techniques or microscopy, are relatively efficient down to low numbers, but there is still a challenge in obtaining sufficiently clean (free of inhibitors) and concentrated samples from the original material to be able to apply the detection methodologies. Thus, efficient removal of the oocysts from food samples using elution solutions, followed by concentration procedures for subsequent detection assays should be areas of focus. In addition, in order to ensure sufficient robustness in terms of sensitivity and specificity, methods developed should be validated in inter-laboratory studies. Collaboration and sharing of material and methods is likely to be of benefit in improving our current analytical techniques.

Keywords

Fresh Produce Polymerase Chain Reaction Detection Cryptosporidium Oocyst Oocyst Wall Oocyst Excretion 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. Aksoy U, Marangi M, Papini R, Ozkoc S, Bayram Delibas S, Giangaspero A (2014) Detection of toxoplasma gondii and cyclospora cayetanensis in Mytilus galloprovincialis from Izmir Province coast (Turkey) by real time PCR/high-resolution melting analysis (HRM). Food Microbiol 44:128–135CrossRefGoogle Scholar
  2. Chandra V, Torres M, Ortega YR (2014) Efficacy of wash solutions in recovering cyclospora cayetanensis, Cryptosporidium parvum, and toxoplasma gondii from basil. J Food Prot 77:1348–1354CrossRefGoogle Scholar
  3. Cinar HN, Gopinath G, Jarvis K, Murphy HR (2015) The complete mitochondrial genome of the foodborne parasitic pathogen cyclospora cayetanensis. PLoS One 10:e0128645CrossRefGoogle Scholar
  4. Di Gliullo AB, Cribari MS, Bava AJ, Cicconetti JS, Collazos R (2000) Cyclospora cayetanensis in sputum and stool samples. Rev Inst Med Trop Sao Paulo 42:115–117CrossRefGoogle Scholar
  5. Eberhard ML, Pieniazek NJ, Arrowood MJ (1997) Laboratory diagnosis of cyclospora infections. Arch Pathol Lab Med 121:792–797Google Scholar
  6. Graczyk TK, Ortega YR, Conn DB (1998) Recovery of waterborne oocysts of cyclospora cayetanensis by Asian freshwater clams (Corbicula fluminea). Am J Trop Med Hyg 59:928–932Google Scholar
  7. Guo Y, Roellig DM, Li N, Tang K, Frace M, Ortega Y et al (2016) Multilocus sequence typing tool for cyclospora cayetanensis. Emerg Infect Dis 22:1464–1467CrossRefGoogle Scholar
  8. Hussein EM (2007) Molecular identification of Cycospora spp. using multiplex PCR from diarrheic children compared to others conventional methods. J Egypt Soc Parasitol 37:585–598Google Scholar
  9. Hussein EM, El-Moamly AA, Dawoud HA, Fahmy H, El-Shal HE, Sabek NA (2007) Real-time PCR and flow cytometry in detection of cyclospora oocysts in fecal samples of symptomatic and asymptomatic pediatrics patients. J Egypt Soc Parasitol 37:151–170Google Scholar
  10. Hussein EM, El-Moamly AA, Mahmoud MA, Ateek NS (2016) Wide genetic variations at 18S ribosomal RNA locus of cyclospora cayetanensis isolated from Egyptian patients using high resolution melting curve. Parasitol Res 115:2797–2806CrossRefGoogle Scholar
  11. Jinneman KC, Wetherington JH, Hill WE, Adams AM, Johnson JM, Tenge BJ et al (1998) Template preparation for PCR and RFLP of amplification products for the detection and identification of cyclospora sp. and Eimeria spp. oocysts directly from raspberries. J Food Prot 61:1497–1503CrossRefGoogle Scholar
  12. Jinneman KC, Wetherington JH, Hill WE, Omiescinski CJ, Adams AM, Johnson JM et al (1999) An oligonucleotide-ligation assay for the differentiation between cyclospora and Eimeria spp. polymerase chain reaction amplification products. J Food Prot 62:682–685CrossRefGoogle Scholar
  13. Lalonde LF, Gajadhar AA (2008) Highly sensitive and specific PCR assay for reliable detection of cyclospora cayetanensis oocysts. Appl Environ Microbiol 74:4354–4358CrossRefGoogle Scholar
  14. Lalonde LF, Gajadhar AA (2011) Detection and differentiation of coccidian oocysts by real-time PCR and melting curve analysis. J Parasitol 97:725–730CrossRefGoogle Scholar
  15. Lalonde LF, Reyes J, Gajadhar AA (2013) Application of a qPCR assay with melting curve analysis for detection and differentiation of protozoan oocysts in human fecal samples from Dominican Republic. Am J Trop Med Hyg 89:892–898CrossRefGoogle Scholar
  16. Lee SH, Joung M, Yoon S, Choi K, Park WY, Yu JR (2010) Multiplex PCR detection of waterborne intestinal protozoa: microsporidia, cyclospora, and cryptosporidium. Korean J Parasitol 48:297–301CrossRefGoogle Scholar
  17. Liu S, Wang L, Zheng H, Xu Z, Roellig DM, Li N et al (2016) Comparative genomics reveals cyclospora cayetanensis possesses coccidia-like metabolism and invasion components but unique surface antigens. BMC Genomics 17:316CrossRefGoogle Scholar
  18. Marangi M, Koehler AV, Zanzani SA, Manfredi MT, Brianti E, Giangaspero A et al (2015) Detection of cyclospora in captive chimpanzees and macaques by a quantitative PCR-based mutation scanning approach. Parasit Vectors 8:274CrossRefGoogle Scholar
  19. Nace EK, Steurer FJ, Eberhard ML (1999) Evaluation of Streck tissue fixative, a nonformalin fixative for preservation of stool samples and subsequent parasitologic examination. J Clin Microbiol 37:4113–4119Google Scholar
  20. Ogedengbe ME, Qvarnstrom Y, da Silva AJ, Arrowood MJ, Barta JR (2015) A linear mitochondrial genome of cyclospora cayetanensis (Eimeriidae, Eucoccidiorida, Coccidiasina, Apicomplexa) suggests the ancestral start position within mitochondrial genomes of eimeriid coccidia. Int J Parasitol 45:361–365CrossRefGoogle Scholar
  21. Orlandi PA, Lampel KA (2000) Extraction-free, filter-based template preparation for rapid and sensitive PCR detection of pathogenic parasitic protozoa. J Clin Microbiol 38:2271–2277Google Scholar
  22. Orlandi PA, Carter L, Brinker AM, da Silva AJ, Chu DM, Lampel KA et al (2003) Targeting single-nucleotide polymorphisms in the 18S rRNA gene to differentiate cyclospora species from Eimeria species by multiplex PCR. Appl Environ Microbiol 69:4806–4813CrossRefGoogle Scholar
  23. Orlandi PA, Frazar C, Carter L, Chu D-M (2004a) BAM: detection of cyclospora and cryptosporidiumGoogle Scholar
  24. Orlandi PA, Frazar C, Carter L, Chu DM (2004b) Chapter 19A. Detection of Cyclospora ad Cryptosporidium from fresh produce: isolation and identification by polymerase chain reaction (PCR) and microscopic analysis. In: Bacteriological analytical manualGoogle Scholar
  25. Ortega YR, Sterling CR, Gilman RH, Cama VA, Diaz F (1993) Cyclospora species—a new protozoan pathogen of humans. N Engl J Med 328:1308–1312CrossRefGoogle Scholar
  26. Ortega YR, Gilman RH, Sterling CR (1994) A new coccidian parasite (Apicomplexa: Eimeriidae) from humans. J Parasitol 80:625–629CrossRefGoogle Scholar
  27. Qvarnstrom Y, Wei-Pridgeon Y, Li W, Nascimento FS, Bishop HS, Herwaldt BL et al (2015) Draft genome sequences from cyclospora cayetanensis oocysts purified from a human stool sample. Genom Announc 3:e01324CrossRefGoogle Scholar
  28. Relman DA, Schmidt TM, Gajadhar A, Sogin M, Cross J, Yoder K et al (1996) Molecular phylogenetic analysis of cyclospora, the human intestinal pathogen, suggests that it is closely related to Eimeria species. J Infect Dis 173:440–445CrossRefGoogle Scholar
  29. Robertson LJ, Gjerde B (2001) Factors affecting recovery efficiency in isolation of cryptosporidium oocysts and giardia cysts from vegetables for standard method development. J Food Prot 64:1799–1805CrossRefGoogle Scholar
  30. Robertson LJ, Gjerde B, Campbell AT (2000) Isolation of cyclospora oocysts from fruits and vegetables using lectin-coated paramagnetic beads. J Food Prot 63:1410–1414CrossRefGoogle Scholar
  31. Shields JM, Olson BH (2003) PCR-restriction fragment length polymorphism method for detection of cyclospora cayetanensis in environmental waters without microscopic confirmation. Appl Environ Microbiol 69:4662–4669CrossRefGoogle Scholar
  32. Shields JM, Lee MM, Murphy HR (2012) Use of a common laboratory glassware detergent improves recovery of Cryptosporidium parvum and cyclospora cayetanensis from lettuce, herbs and raspberries. Int J Food Microbiol 153:123–128CrossRefGoogle Scholar
  33. Sulaiman IM, Torres P, Simpson S, Kerdahi K, Ortega Y (2013) Sequence characterization of heat shock protein gene of cyclospora cayetanensis isolates from Nepal, Mexico, and Peru. J Parasitol 99:379–382CrossRefGoogle Scholar
  34. Sulaiman IM, Ortega Y, Simpson S, Kerdahi K (2014) Genetic characterization of human-pathogenic cyclospora cayetanensis parasites from three endemic regions at the 18S ribosomal RNA locus. Infect Genet Evol 22:229–234CrossRefGoogle Scholar
  35. Tang K, Guo Y, Zhang L, Rowe LA, Roellig DM, Frace MA et al (2015) Genetic similarities between cyclospora cayetanensis and cecum-infecting avian Eimeria spp. in apicoplast and mitochondrial genomes. Parasit Vectors 8:358CrossRefGoogle Scholar
  36. Varma M, Hester JD, Schaefer FW 3rd, Ware MW, Lindquist HD (2003) Detection of cyclospora cayetanensis using a quantitative real-time PCR assay. J Microbiol Methods 53:27–36CrossRefGoogle Scholar
  37. Visvesvara GS, Moura H, Kovacs-Nace E, Wallace S, Eberhard ML (1997) Uniform staining of cyclospora oocysts in fecal smears by a modified safranin technique with microwave heating. J Clin Microbiol 35:730–733Google Scholar
  38. Ye J, Xiao L, Li J, Huang W, Amer SE, Guo Y et al (2014) Occurrence of human-pathogenic Enterocytozoon bieneusi, giardia duodenalis and cryptosporidium genotypes in laboratory macaques in Guangxi. China Parasitol Int 63:132–137CrossRefGoogle Scholar

Copyright information

© The Author(s) 2017

Authors and Affiliations

  • Ynés R. Ortega
    • 1
  • Lucy J. Robertson
    • 2
  1. 1.Center for Food SafetyUniversity of GeorgiaGriffinUSA
  2. 2.Parasitology, Faculty of Veterinary MedicineNorwegian University of Life SciencesOsloNorway

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