Research Article
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Investigation of Cryptosporidium spp. in Immunosuppressive and Immunocompetent Cases with Diarrhea by Microscopic, Serological and Molecular Methods

Year 2018, Volume: 4 Issue: 3, 11 - 19, 27.12.2018
https://doi.org/10.19127/mbsjohs.464833

Abstract

Objective: In this study, our aim
was to compare the diagnostic methods with each other and show the presence of Cryptosporidium oocysts by using
molecular, serological and microscopic methods in stool samples which were collected
from immunosuppressive and immunocompetent patients with diarrhea

Methods: Total 172 stool samples were
collected from 80 immunosuppressive patients and 92 immunocompetent patients
(between 0-94 years) with diarrhea. These stool samples were obtained from the
different clinics of Ataturk University, Yakutiye Research Hospital between
January 2014 and July 2014. Patient group composed of 49 persons between 0-14
years and 123 persons between 15-94 years. On the other hand, 141 patients were
using tap water while 41 of them were using well water. Modified acid-fast
staining, ELISA and DFA techniques were applied to detect the Cryptosporidium
parasite positivity. Nested PCR method was performed to the samples which were
detected positive with one of the above methods.

Results: The positivity was detected in
5.8%, 4.1% and 3.5% by ELISA, DFA technique and Modified acid-fast staining
method, respectively.  Cryptosporidium
DNA was detected in only 1.2% by PCR method. The rates of positivity were 6.3%
and 5.4% in immunosuppressive and immunocompetent patients, respectively. The
positivity was detected in 10.2% and 4.1% in 0-14 age group and 15-94 age group
patients, respectively. On the other hand, 4.3% and 12.9% positivity rates were
detected in tap water and well water users respectively.







Conclusion: Our study pointed out that the investigation of
Cryptosporidium oocysts as diarrhea agents in especially immunosuppressive
patients, individuals in childhood and well water users may be useful. Because
cryptosporidiosis is a common disease in children and immunosuppressive
individuals. Additionally, we think that ELISA method can be preferred to other
methods in terms of high sensitivity and ease of application.

References

  • Alabdeen, A.Z. 2014. 'İshalli Hastalarda Cryptosporidiums spp'nin Modifiye Ehrlich-Ziehl-Neelsen Boyama Yöntemi ve İmmünkromatografik Yöntem ile Araştırılması', Yüksek Lisans, Gaziantep Üniversitesi.
  • Alves, M., L. Xiao, F. Antunes, and O. Matos. 2006. 'Distribution of Cryptosporidium subtypes in humans and domestic and wild ruminants in Portugal', Parasitol Res, 99: 287-92.
  • Amar, C. F., P. H. Dear, and J. McLauchlin. 2004. 'Detection and identification by real time PCR/RFLP analyses of Cryptosporidium species from human faeces', Lett Appl Microbiol, 38: 217-22.
  • Baumgartner, A., H. P. Marder, J. Munzinger, and H. H. Siegrist. 2000. 'Frequency of Cryptosporidium spp. as cause of human gastrointestinal disease in Switzerland and possible sources of infection', Schweiz Med Wochenschr, 130: 1252-8.
  • Bera, P., S. Das, R. Saha, V. G. Ramachandran, and D. Shah. 2014. 'Cryptosporidium in children with diarrhea: a hospital-based Study', Indian Pediatr, 51: 906-8.
  • Botero, J. H., A. Castano, M. N. Montoya, N. E. Ocampo, M. I. Hurtado, and M. M. Lopera. 2003. 'A preliminary study of the prevalence of intestinal parasites in immunocompromised patients with and without gastrointestinal manifestations', Rev Inst Med Trop Sao Paulo, 45: 197-200.
  • Clark, D. P. 1999. 'New insights into human cryptosporidiosis', Clin Microbiol Rev, 12: 554-63.
  • Ekinci, A.İ. 2012. 'Kars Yöresinde Sığırlarda Cryptosporidium Türlerinin Dağılımı ve Moleküler Karekterizasyonu', Doktora Tezi, Kafkas Üniversitesi.
  • Elgün, G. 2009. 'İshalli Dışkı Örneklerinde Cryptosporidium Sp. Antijenin ELİSA Yöntemi ile Araştırılması', Yüksek Lisans, Çukurova Üniversitesi.
  • Eren, C. 2011. 'Farklı Gruplardaki İmmün-Süprese Bireylerde Cryptosporidium'un ELİSA ve Modifiye Asit-Fast Boyama Yöntemi ile Araştırılması', Uzmanlık Tezi, Dicle Üniversitesi.
  • Fathy, M. M., N. M. Abdelrazek, F. A. Hassan, and A. A. El-Badry. 2014. 'Molecular copro-prevalence of Cryptosporidium in Egyptian children and evaluation of three diagnostic methods', Indian Pediatr, 51: 727-9.
  • Fayer, R. 2004. 'Cryptosporidium: a water-borne zoonotic parasite', Vet Parasitol, 126: 37-56.
  • Fayer. 2010. 'Taxonomy and species delimitation in Cryptosporidium', Exp Parasitol, 124: 90-7.
  • Fayer, R., U. Morgan, and S. J. Upton. 2000. 'Epidemiology of Cryptosporidium: transmission, detection and identification', Int J Parasitol, 30: 1305-22.
  • Galvan, A. L., A. Magnet, F. Izquierdo, C. Fernandez Vadillo, R. H. Peralta, S. Angulo, S. Fenoy, and C. del Aguila. 2014. 'A year-long study of Cryptosporidium species and subtypes in recreational, drinking and wastewater from the central area of Spain', Sci Total Environ, 468-469: 368-75.
  • Garcia, L. S., D. A. Bruckner, T. C. Brewer, and R. Y. Shimizu. 1983. 'Techniques for the recovery and identification of Cryptosporidium oocysts from stool specimens', J Clin Microbiol, 18: 185-90.
  • Gatei, W., C. N. Wamae, C. Mbae, A. Waruru, E. Mulinge, T. Waithera, S. M. Gatika, S. K. Kamwati, G. Revathi, and C. A. Hart. 2006. 'Cryptosporidiosis: prevalence, genotype analysis, and symptoms associated with infections in children in Kenya', Am J Trop Med Hyg, 75: 78-82.
  • Hunter, P. R., S. Hughes, S. Woodhouse, Q. Syed, N. Q. Verlander, R. M. Chalmers, K. Morgan, G. Nichols, N. Beeching, and K. Osborn. 2004. 'Sporadic cryptosporidiosis case-control study with genotyping', Emerg Infect Dis, 10: 1241-9.
  • Jayalakshmi, J., B. Appalaraju, and K. Mahadevan. 2008. 'Evaluation of an enzyme-linked immunoassay for the detection of Cryptosporidium antigen in fecal specimens of HIV/AIDS patients', Indian J Pathol Microbiol, 51: 137-8.
  • Kar, S. 2007. 'Cryptosporidium parvum'un Hücre Kültüründe Üretilmesi ve Üremenin Farklı Yöntemlerle Belirlenmesi', Doktora Tezi, Ankara Üniversitesi.Kehl, K. S., H. Cicirello, and P. L. Havens. 1995. 'Comparison of four different methods for detection of Cryptosporidium species', J Clin Microbiol, 33: 416-8.
  • Koturoglu, G., S. Bayram, Z. Kurugol, N. Turgay, and F. Mutlubas. 2004. 'Akut ishalli çocuklarda Cryptosporidium sıklığı ve risk faktörleri', T Klin J Pediatr, 13: 1-19.Leng, X., D. A. Mosier, and R. D. Oberst. 1996. 'Simplified method for recovery and PCR detection of Cryptosporidium DNA from bovine feces', Appl Environ Microbiol, 62: 643-7.
  • MacPherson, D. W., and R. McQueen. 1993. 'Cryptosporidiosis: multiattribute evaluation of six diagnostic methods', J Clin Microbiol, 31: 198-202.
  • Magi, B., V. Canocchi, G. Tordini, C. Cellesi, and A. Barberi. 2006. 'Cryptosporidium infection: diagnostic techniques', Parasitol Res, 98: 150-2.
  • Morgan, U. M., L. Pallant, B. W. Dwyer, D. A. Forbes, G. Rich, and R. C. Thompson. 1998. 'Comparison of PCR and microscopy for detection of Cryptosporidium parvum in human fecal specimens: clinical trial', J Clin Microbiol, 36: 995-8.
  • Moss, D. M., S. N. Bennett, M. J. Arrowood, S. P. Wahlquist, and P. J. Lammie. 1998. 'Enzyme-linked immunoelectrotransfer blot analysis of a cryptosporidiosis outbreak on a United States Coast Guard cutter', Am J Trop Med Hyg, 58: 110-8.
  • Omoruyi, B. E., U. U. Nwodo, C. S. Udem, and F. O. Okonkwo. 2014. 'Comparative diagnostic techniques for cryptosporidium infection', Molecules, 19: 2674-83.
  • Pereira, M. D., E. R. Atwill, A. P. Barbosa, S. A. Silva, and M. T. Garcia-Zapata. 2002. 'Intra-familial and extra-familial risk factors associated with Cryptosporidium parvum infection among children hospitalized for diarrhea in Goiania, Goias, Brazil', Am J Trop Med Hyg, 66: 787-93.
  • Ramratnam, B., and T. P. Flanigan. 1997. 'Cryptosporidiosis in persons with HIV infection', Postgrad Med J, 73: 713-6.
  • Rosenblatt, J. E., and L. M. Sloan. 1993. 'Evaluation of an enzyme-linked immunosorbent assay for detection of Cryptosporidium spp. in stool specimens', J Clin Microbiol, 31: 1468-71.
  • Sears, C. L., and B.D. Kirckpatrick. 2001. Cryptosporidiosis and İsosporidiosis. Principles and Practice of Clinical Parasitology. (John Wiley & Sons Ltd. Pres.).
  • Sirisena, U. M., W. M. Iddawela, F. Noordeen, and S. Wickramasinghe. 2014. 'Prevalence and identification of Cryptosporidium species in paediatric patients with diarrhoea', Ceylon Med J, 59: 75-8.
  • Sreedharan, A., R. S. Jayshree, and H. Sridhar. 1996. 'Cryptosporidiosis among cancer patients: an observation', J Diarrhoeal Dis Res, 14: 211-3.Starling, C.R., and M.J. Arrowood. 1993. Cryptosporidia, İn: Parazitic Protozoa (Academic Press).
  • Tanyuksel, M., H. Gun, and L. Doganci. 1995. 'Prevalence of Cryptosporidium sp. in patients with neoplasia and diarrhea', Scand J Infect Dis, 27: 69-70.Usluca, S. 2009. 'İshalli Dışkılarda Microsporidium Spp. ve Cryptosporidium Spp.'nin Saptanması PCR Yöntemi ile Tür Tayininin Yapılması', Doktora Tezi, Dokuz Eylül Üniversitesi.
  • Xiao, L., C. Bern, J. Limor, I. Sulaiman, J. Roberts, W. Checkley, L. Cabrera, R. H. Gilman, and A. A. Lal. 2001. 'Identification of 5 types of Cryptosporidium parasites in children in Lima, Peru', J Infect Dis, 183: 492-7.
  • Xiao, L., R. Fayer, U. Ryan, and S. J. Upton. 2004. 'Cryptosporidium taxonomy: recent advances and implications for public health', Clin Microbiol Rev, 17: 72-97.
  • Xiao, L., I. M. Sulaiman, U. M. Ryan, L. Zhou, E. R. Atwill, M. L. Tischler, X. Zhang, R. Fayer, and A. A. Lal. 2002. 'Host adaptation and host-parasite co-evolution in Cryptosporidium: implications for taxonomy and public health', Int J Parasitol, 32: 1773-85.
  • Yilmaz, H., Z. Tas-Cengiz, and M. Cicek. 2008. 'Investigation of cryptosporidiosis by enzyme-linked immunosorbent assay and microscopy in children with diarrhea', Saudi Med J, 29: 526-9.
Year 2018, Volume: 4 Issue: 3, 11 - 19, 27.12.2018
https://doi.org/10.19127/mbsjohs.464833

Abstract

References

  • Alabdeen, A.Z. 2014. 'İshalli Hastalarda Cryptosporidiums spp'nin Modifiye Ehrlich-Ziehl-Neelsen Boyama Yöntemi ve İmmünkromatografik Yöntem ile Araştırılması', Yüksek Lisans, Gaziantep Üniversitesi.
  • Alves, M., L. Xiao, F. Antunes, and O. Matos. 2006. 'Distribution of Cryptosporidium subtypes in humans and domestic and wild ruminants in Portugal', Parasitol Res, 99: 287-92.
  • Amar, C. F., P. H. Dear, and J. McLauchlin. 2004. 'Detection and identification by real time PCR/RFLP analyses of Cryptosporidium species from human faeces', Lett Appl Microbiol, 38: 217-22.
  • Baumgartner, A., H. P. Marder, J. Munzinger, and H. H. Siegrist. 2000. 'Frequency of Cryptosporidium spp. as cause of human gastrointestinal disease in Switzerland and possible sources of infection', Schweiz Med Wochenschr, 130: 1252-8.
  • Bera, P., S. Das, R. Saha, V. G. Ramachandran, and D. Shah. 2014. 'Cryptosporidium in children with diarrhea: a hospital-based Study', Indian Pediatr, 51: 906-8.
  • Botero, J. H., A. Castano, M. N. Montoya, N. E. Ocampo, M. I. Hurtado, and M. M. Lopera. 2003. 'A preliminary study of the prevalence of intestinal parasites in immunocompromised patients with and without gastrointestinal manifestations', Rev Inst Med Trop Sao Paulo, 45: 197-200.
  • Clark, D. P. 1999. 'New insights into human cryptosporidiosis', Clin Microbiol Rev, 12: 554-63.
  • Ekinci, A.İ. 2012. 'Kars Yöresinde Sığırlarda Cryptosporidium Türlerinin Dağılımı ve Moleküler Karekterizasyonu', Doktora Tezi, Kafkas Üniversitesi.
  • Elgün, G. 2009. 'İshalli Dışkı Örneklerinde Cryptosporidium Sp. Antijenin ELİSA Yöntemi ile Araştırılması', Yüksek Lisans, Çukurova Üniversitesi.
  • Eren, C. 2011. 'Farklı Gruplardaki İmmün-Süprese Bireylerde Cryptosporidium'un ELİSA ve Modifiye Asit-Fast Boyama Yöntemi ile Araştırılması', Uzmanlık Tezi, Dicle Üniversitesi.
  • Fathy, M. M., N. M. Abdelrazek, F. A. Hassan, and A. A. El-Badry. 2014. 'Molecular copro-prevalence of Cryptosporidium in Egyptian children and evaluation of three diagnostic methods', Indian Pediatr, 51: 727-9.
  • Fayer, R. 2004. 'Cryptosporidium: a water-borne zoonotic parasite', Vet Parasitol, 126: 37-56.
  • Fayer. 2010. 'Taxonomy and species delimitation in Cryptosporidium', Exp Parasitol, 124: 90-7.
  • Fayer, R., U. Morgan, and S. J. Upton. 2000. 'Epidemiology of Cryptosporidium: transmission, detection and identification', Int J Parasitol, 30: 1305-22.
  • Galvan, A. L., A. Magnet, F. Izquierdo, C. Fernandez Vadillo, R. H. Peralta, S. Angulo, S. Fenoy, and C. del Aguila. 2014. 'A year-long study of Cryptosporidium species and subtypes in recreational, drinking and wastewater from the central area of Spain', Sci Total Environ, 468-469: 368-75.
  • Garcia, L. S., D. A. Bruckner, T. C. Brewer, and R. Y. Shimizu. 1983. 'Techniques for the recovery and identification of Cryptosporidium oocysts from stool specimens', J Clin Microbiol, 18: 185-90.
  • Gatei, W., C. N. Wamae, C. Mbae, A. Waruru, E. Mulinge, T. Waithera, S. M. Gatika, S. K. Kamwati, G. Revathi, and C. A. Hart. 2006. 'Cryptosporidiosis: prevalence, genotype analysis, and symptoms associated with infections in children in Kenya', Am J Trop Med Hyg, 75: 78-82.
  • Hunter, P. R., S. Hughes, S. Woodhouse, Q. Syed, N. Q. Verlander, R. M. Chalmers, K. Morgan, G. Nichols, N. Beeching, and K. Osborn. 2004. 'Sporadic cryptosporidiosis case-control study with genotyping', Emerg Infect Dis, 10: 1241-9.
  • Jayalakshmi, J., B. Appalaraju, and K. Mahadevan. 2008. 'Evaluation of an enzyme-linked immunoassay for the detection of Cryptosporidium antigen in fecal specimens of HIV/AIDS patients', Indian J Pathol Microbiol, 51: 137-8.
  • Kar, S. 2007. 'Cryptosporidium parvum'un Hücre Kültüründe Üretilmesi ve Üremenin Farklı Yöntemlerle Belirlenmesi', Doktora Tezi, Ankara Üniversitesi.Kehl, K. S., H. Cicirello, and P. L. Havens. 1995. 'Comparison of four different methods for detection of Cryptosporidium species', J Clin Microbiol, 33: 416-8.
  • Koturoglu, G., S. Bayram, Z. Kurugol, N. Turgay, and F. Mutlubas. 2004. 'Akut ishalli çocuklarda Cryptosporidium sıklığı ve risk faktörleri', T Klin J Pediatr, 13: 1-19.Leng, X., D. A. Mosier, and R. D. Oberst. 1996. 'Simplified method for recovery and PCR detection of Cryptosporidium DNA from bovine feces', Appl Environ Microbiol, 62: 643-7.
  • MacPherson, D. W., and R. McQueen. 1993. 'Cryptosporidiosis: multiattribute evaluation of six diagnostic methods', J Clin Microbiol, 31: 198-202.
  • Magi, B., V. Canocchi, G. Tordini, C. Cellesi, and A. Barberi. 2006. 'Cryptosporidium infection: diagnostic techniques', Parasitol Res, 98: 150-2.
  • Morgan, U. M., L. Pallant, B. W. Dwyer, D. A. Forbes, G. Rich, and R. C. Thompson. 1998. 'Comparison of PCR and microscopy for detection of Cryptosporidium parvum in human fecal specimens: clinical trial', J Clin Microbiol, 36: 995-8.
  • Moss, D. M., S. N. Bennett, M. J. Arrowood, S. P. Wahlquist, and P. J. Lammie. 1998. 'Enzyme-linked immunoelectrotransfer blot analysis of a cryptosporidiosis outbreak on a United States Coast Guard cutter', Am J Trop Med Hyg, 58: 110-8.
  • Omoruyi, B. E., U. U. Nwodo, C. S. Udem, and F. O. Okonkwo. 2014. 'Comparative diagnostic techniques for cryptosporidium infection', Molecules, 19: 2674-83.
  • Pereira, M. D., E. R. Atwill, A. P. Barbosa, S. A. Silva, and M. T. Garcia-Zapata. 2002. 'Intra-familial and extra-familial risk factors associated with Cryptosporidium parvum infection among children hospitalized for diarrhea in Goiania, Goias, Brazil', Am J Trop Med Hyg, 66: 787-93.
  • Ramratnam, B., and T. P. Flanigan. 1997. 'Cryptosporidiosis in persons with HIV infection', Postgrad Med J, 73: 713-6.
  • Rosenblatt, J. E., and L. M. Sloan. 1993. 'Evaluation of an enzyme-linked immunosorbent assay for detection of Cryptosporidium spp. in stool specimens', J Clin Microbiol, 31: 1468-71.
  • Sears, C. L., and B.D. Kirckpatrick. 2001. Cryptosporidiosis and İsosporidiosis. Principles and Practice of Clinical Parasitology. (John Wiley & Sons Ltd. Pres.).
  • Sirisena, U. M., W. M. Iddawela, F. Noordeen, and S. Wickramasinghe. 2014. 'Prevalence and identification of Cryptosporidium species in paediatric patients with diarrhoea', Ceylon Med J, 59: 75-8.
  • Sreedharan, A., R. S. Jayshree, and H. Sridhar. 1996. 'Cryptosporidiosis among cancer patients: an observation', J Diarrhoeal Dis Res, 14: 211-3.Starling, C.R., and M.J. Arrowood. 1993. Cryptosporidia, İn: Parazitic Protozoa (Academic Press).
  • Tanyuksel, M., H. Gun, and L. Doganci. 1995. 'Prevalence of Cryptosporidium sp. in patients with neoplasia and diarrhea', Scand J Infect Dis, 27: 69-70.Usluca, S. 2009. 'İshalli Dışkılarda Microsporidium Spp. ve Cryptosporidium Spp.'nin Saptanması PCR Yöntemi ile Tür Tayininin Yapılması', Doktora Tezi, Dokuz Eylül Üniversitesi.
  • Xiao, L., C. Bern, J. Limor, I. Sulaiman, J. Roberts, W. Checkley, L. Cabrera, R. H. Gilman, and A. A. Lal. 2001. 'Identification of 5 types of Cryptosporidium parasites in children in Lima, Peru', J Infect Dis, 183: 492-7.
  • Xiao, L., R. Fayer, U. Ryan, and S. J. Upton. 2004. 'Cryptosporidium taxonomy: recent advances and implications for public health', Clin Microbiol Rev, 17: 72-97.
  • Xiao, L., I. M. Sulaiman, U. M. Ryan, L. Zhou, E. R. Atwill, M. L. Tischler, X. Zhang, R. Fayer, and A. A. Lal. 2002. 'Host adaptation and host-parasite co-evolution in Cryptosporidium: implications for taxonomy and public health', Int J Parasitol, 32: 1773-85.
  • Yilmaz, H., Z. Tas-Cengiz, and M. Cicek. 2008. 'Investigation of cryptosporidiosis by enzyme-linked immunosorbent assay and microscopy in children with diarrhea', Saudi Med J, 29: 526-9.
There are 37 citations in total.

Details

Primary Language English
Subjects Health Care Administration
Journal Section Research articles
Authors

Ahmet Yılmaz

Önder Akkaş This is me

Esin Güven

Hakan Aydın

Hakan Uslu This is me

Publication Date December 27, 2018
Published in Issue Year 2018 Volume: 4 Issue: 3

Cite

Vancouver Yılmaz A, Akkaş Ö, Güven E, Aydın H, Uslu H. Investigation of Cryptosporidium spp. in Immunosuppressive and Immunocompetent Cases with Diarrhea by Microscopic, Serological and Molecular Methods. Mid Blac Sea J Health Sci. 2018;4(3):11-9.

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