Research Article
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Year 2019, , 250 - 255, 30.06.2019
https://doi.org/10.7197/223.vi.476804

Abstract

References

  • 1. Hashwey R, Smith NH, Cron S, Graviss EA, Chappell, CL, White AC. Cryptosporidiosis in Houston. Texas a report of 95 casesMedicine1997; 76(2): 118-139.
  • 2. Özçelik S, Poyraz Ö, Kalkan K, Malatyalı E, Değerli S. Hayvancılıkla uğraşanlarda ve sığırlardaCryptosporidiumspp. Yaygınlığının ELISA ile araştırılması. Kafkas UnivVet Fak Derg2012;18:A61-A64, DOI:10.9775/kvfd.2011.5981.
  • 3. Clark DP. New insightsintohumancryptosporidiosis. ClinMicrobiolRev 1999; 12:554–563.
  • 4. Griffiths JK. Human cryptosporidiosis: epidemiology, transmission, clinicaldisease, treatment, anddiagnosis. AdvParasitol 1998; 40:37–85.
  • 5. Miron D, Kenes J, Dagan R. Calves as a source of an outbreak of cryptosporidiosisamongyoungchildren in an agriculturalclosedcommunity. Pediatr InfectDis J 1991; 10:438–441.
  • 6. Çeliksöz A, Çelik S. Cumhuriyet Üniversitesi Hastanesi’nde gastroenteritli ve malnütrisyonlu hastalarda Cryptosporidiumspp. Araştırması. Türkiye ParazitolDerg 2003; 27: 85-88.
  • 7. Egyed Z, Sreter T, Szell Z, Varga L. Characterization of Cryptosporidiumspp.- recentdevelopmentsandfutureneeds. Vet. Parasitol 2003; 111:103-114.
  • 8. Usluca S. İshalli Dışkılarda Microsporidiumspp. veCryptosporidiumspp.’ nin Saptanması PCR Yöntemi İle Tür Tayininin Yapılması. Doktora tezi. 2009.
  • 9. Börekçi G, Otağ F, Emekdaş G. Mersin’de bir gecekondu mahallesinde yaşayan ailelerde Cryptosporidiumprevalansı. İnfekDerg 2005;19(1):39-46.
  • 10. Tanyüksel M, Haznedaroğlu T, Gün H. Neoplastik hastalarda Cryptosporidiumspp. Araştırılması. Türkiye ParazitolDerg1995; 19 (1): 56-63.
  • 11. Hazer Y. Afyonkarahisar bölgesindeki risk gruplarında Cryptosporidiumparvum araştırılması. Afyonkarahisar:Afyonkarahisar Kocatepe Üniversitesi Sağlık Bilimleri Enstitüsü. 2007.
  • 12. Karanis P, Kimura A. Evaluation of three flocculation methods for the purification of Cryptosporidium parvum oocysts from water samples. Lett ApplMicrobiol2002; 34:444–449.
  • 13. Plutzer J. CryptosporidiumandGiardiaas watercontaminantpathogens in Hungary: 2008; 22-25.
  • 14. Dağ A. Şanlıurfa yöresinde immunsuprese hastalarda Cryptosporidiumspp. sıklığınınkinyoun asit-fast boyama ve elisa yöntemleri ile araştırılması.Uzmanlık Tezi, Harran Üniversitesi, Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, Şanlıurfa. 2010.
  • 15. Karaman U, Dakdal N, Özer A, Enginyurt Ö, Ertürk Ö. Incıdence of Cryptosporidiumspp. Inthehumanpopulatıon of malatya in Turkey, ActaMedMediterr2015, 31: 263.
  • 16. Yılmaz, H., Taş, C.Z., Çiçek, M. Investigation of cryptosporidiosisbyenzyme-linkedimmunosorbentassayandmicroscopy in childrenwithdiarrhea. SaudiMed J 2008; 29(4): 526-529.
  • 17. Elgün, G.,Koltaş, I.S. Investigation of Cryptosporidiumspp. antigenby ELISA method in stoolspecimensobtainedfrompatientswithdiarrhea. ParasitolRes2011;108(2): 395-7.
  • 18. Sungur T, Kar S, Güven E, Aktaş M, Karaer Z, Vatansever Z. Cryptosporidiumspp’nin dışkıdan Nested PCR ve carbolfuchsin boyama yöntemi ile teşhis edilmesi. Türkiye ParazitolDerg 2008; 32(4): 305-308.
  • 19. Sakarya Y, Kar S, Tanyuksel M. ve ark. Detection of Cryptosporidiumspp. inhumansandcalvesthroughNested PCR andcarbolfuchsinstainingmethods in Ankara, Turkey. Kafkas UnivVet Fak Derg2010; 977-980.
  • 20. Mojarad, N.E.,Keshavarz, A., Taghipour, N., Haghighi, A., Kazemi, B., Athari, A. Genotyping of Cryptosporidiumspp. inclinicalsamples: PCR-RFLP analysis of the TRAP-C2 gene. GastroenterolHepatolBedBench2011; 4(1): 29-33.
  • 21. Ülçay A., Görenek L., Coşkun Ö., Araz E., Acar A., Eyigün, C.P. İmmün yetmezlikli hastalarda intestinal protozoonların tanısı. Türkiye Parazitol Derg 2008; 32(4): 328–333.
  • 22. Usluca, A., Aksoy, Ü. Detection and genotyping of Cryptosporidium spp. in diarrheic stools by PCR/RFLP analyses. Turk J Med Sci 2011; 41(6): 1029-1036.
  • 23. Dirim, E.D., Dağcı, H., Turgay, N., Akarca, U.S., Alkan, M.Z. The molecular diagnosis of cryptosporidiosis in fresh and formalin preserved fecal samples. Türkiye Parazitol Derg 2009; 33(2): 120-4.

Investigation of Cryptosporidium spp. positivity in formol-fixed stool samples by polymerase chain reaction method

Year 2019, , 250 - 255, 30.06.2019
https://doi.org/10.7197/223.vi.476804

Abstract

Objective: It was aimed to determine Cryptosporidium spp., which is thought to cause digestive system complaints
in primary school children, by Modified Acid-Fast (MAF) staining method and
Polymerase Chain Reaction (PCR).

Method: One hundred sixty four stool samples taken from children
were examined under light microscopy with MAF. Positive samples were fixed in
10% formalin and stored at + 4 °C until analysis. These samples were then
purified using the Sucrose Gradient method and, DNA isolation was performed
using the modified QIAamp DNA Mini Kit (Qiagen) protocol. In order to increase
sensitivity of the study, SSU rRNA gene region was amplified by PCR method.

Results: Microscopic examination showed that 89 of the 164 stool
samples (54,26%) were found to be positive for Cryptosporidium spp.oocysts. Eighty nine samples which were found
to be positive by microscopic examination and fixed in %10 formalin were
studied by PCR and 74 (45,12%) of them were positive.







Conclusions: Cryptosporidiosis, which is known to
cause severe infections, especially in immunocompromised individuals and
children, is transmitted by fecal-oral route from infected humans and animals,
and contaminated water and nutrients. Cryptosporidiosis, which is the result of
fecal pollution, causes a great risk for human health even with a low number of
oocysts due to the fact that oocysts are resistant to chlorine and they can
survive for a long time in the outer environment. It was observed that, primary
school children living in area of the study were under risk for
cryptosporidiosis.

References

  • 1. Hashwey R, Smith NH, Cron S, Graviss EA, Chappell, CL, White AC. Cryptosporidiosis in Houston. Texas a report of 95 casesMedicine1997; 76(2): 118-139.
  • 2. Özçelik S, Poyraz Ö, Kalkan K, Malatyalı E, Değerli S. Hayvancılıkla uğraşanlarda ve sığırlardaCryptosporidiumspp. Yaygınlığının ELISA ile araştırılması. Kafkas UnivVet Fak Derg2012;18:A61-A64, DOI:10.9775/kvfd.2011.5981.
  • 3. Clark DP. New insightsintohumancryptosporidiosis. ClinMicrobiolRev 1999; 12:554–563.
  • 4. Griffiths JK. Human cryptosporidiosis: epidemiology, transmission, clinicaldisease, treatment, anddiagnosis. AdvParasitol 1998; 40:37–85.
  • 5. Miron D, Kenes J, Dagan R. Calves as a source of an outbreak of cryptosporidiosisamongyoungchildren in an agriculturalclosedcommunity. Pediatr InfectDis J 1991; 10:438–441.
  • 6. Çeliksöz A, Çelik S. Cumhuriyet Üniversitesi Hastanesi’nde gastroenteritli ve malnütrisyonlu hastalarda Cryptosporidiumspp. Araştırması. Türkiye ParazitolDerg 2003; 27: 85-88.
  • 7. Egyed Z, Sreter T, Szell Z, Varga L. Characterization of Cryptosporidiumspp.- recentdevelopmentsandfutureneeds. Vet. Parasitol 2003; 111:103-114.
  • 8. Usluca S. İshalli Dışkılarda Microsporidiumspp. veCryptosporidiumspp.’ nin Saptanması PCR Yöntemi İle Tür Tayininin Yapılması. Doktora tezi. 2009.
  • 9. Börekçi G, Otağ F, Emekdaş G. Mersin’de bir gecekondu mahallesinde yaşayan ailelerde Cryptosporidiumprevalansı. İnfekDerg 2005;19(1):39-46.
  • 10. Tanyüksel M, Haznedaroğlu T, Gün H. Neoplastik hastalarda Cryptosporidiumspp. Araştırılması. Türkiye ParazitolDerg1995; 19 (1): 56-63.
  • 11. Hazer Y. Afyonkarahisar bölgesindeki risk gruplarında Cryptosporidiumparvum araştırılması. Afyonkarahisar:Afyonkarahisar Kocatepe Üniversitesi Sağlık Bilimleri Enstitüsü. 2007.
  • 12. Karanis P, Kimura A. Evaluation of three flocculation methods for the purification of Cryptosporidium parvum oocysts from water samples. Lett ApplMicrobiol2002; 34:444–449.
  • 13. Plutzer J. CryptosporidiumandGiardiaas watercontaminantpathogens in Hungary: 2008; 22-25.
  • 14. Dağ A. Şanlıurfa yöresinde immunsuprese hastalarda Cryptosporidiumspp. sıklığınınkinyoun asit-fast boyama ve elisa yöntemleri ile araştırılması.Uzmanlık Tezi, Harran Üniversitesi, Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, Şanlıurfa. 2010.
  • 15. Karaman U, Dakdal N, Özer A, Enginyurt Ö, Ertürk Ö. Incıdence of Cryptosporidiumspp. Inthehumanpopulatıon of malatya in Turkey, ActaMedMediterr2015, 31: 263.
  • 16. Yılmaz, H., Taş, C.Z., Çiçek, M. Investigation of cryptosporidiosisbyenzyme-linkedimmunosorbentassayandmicroscopy in childrenwithdiarrhea. SaudiMed J 2008; 29(4): 526-529.
  • 17. Elgün, G.,Koltaş, I.S. Investigation of Cryptosporidiumspp. antigenby ELISA method in stoolspecimensobtainedfrompatientswithdiarrhea. ParasitolRes2011;108(2): 395-7.
  • 18. Sungur T, Kar S, Güven E, Aktaş M, Karaer Z, Vatansever Z. Cryptosporidiumspp’nin dışkıdan Nested PCR ve carbolfuchsin boyama yöntemi ile teşhis edilmesi. Türkiye ParazitolDerg 2008; 32(4): 305-308.
  • 19. Sakarya Y, Kar S, Tanyuksel M. ve ark. Detection of Cryptosporidiumspp. inhumansandcalvesthroughNested PCR andcarbolfuchsinstainingmethods in Ankara, Turkey. Kafkas UnivVet Fak Derg2010; 977-980.
  • 20. Mojarad, N.E.,Keshavarz, A., Taghipour, N., Haghighi, A., Kazemi, B., Athari, A. Genotyping of Cryptosporidiumspp. inclinicalsamples: PCR-RFLP analysis of the TRAP-C2 gene. GastroenterolHepatolBedBench2011; 4(1): 29-33.
  • 21. Ülçay A., Görenek L., Coşkun Ö., Araz E., Acar A., Eyigün, C.P. İmmün yetmezlikli hastalarda intestinal protozoonların tanısı. Türkiye Parazitol Derg 2008; 32(4): 328–333.
  • 22. Usluca, A., Aksoy, Ü. Detection and genotyping of Cryptosporidium spp. in diarrheic stools by PCR/RFLP analyses. Turk J Med Sci 2011; 41(6): 1029-1036.
  • 23. Dirim, E.D., Dağcı, H., Turgay, N., Akarca, U.S., Alkan, M.Z. The molecular diagnosis of cryptosporidiosis in fresh and formalin preserved fecal samples. Türkiye Parazitol Derg 2009; 33(2): 120-4.
There are 23 citations in total.

Details

Primary Language English
Subjects Health Care Administration
Journal Section Basic Science Research Articles
Authors

Ülkü Karaman

Zeynep Kolören

Emine Ayaz

Publication Date June 30, 2019
Acceptance Date May 16, 2019
Published in Issue Year 2019

Cite

AMA Karaman Ü, Kolören Z, Ayaz E. Investigation of Cryptosporidium spp. positivity in formol-fixed stool samples by polymerase chain reaction method. CMJ. June 2019;41(2):250-255. doi:10.7197/223.vi.476804