Abstract
Aim. Cryptosporidium spp. and Giardia intestinalis are important parasitic protozoan causing diarrhea in developing and developed countries. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive and time-consuming. Thus, we aimed the presence of these protozoa in patients with diarrhea and to evaluate the usefulness of direct fluoresan antibody (DFA) test in detecting Cryptosporidium spp and G.intestinalis. from fecal specimens. Methods. For this aim, microscopy and specific antigen detection methods were used to determine Cryptosporidium spp. and G.intestinalis. One hundred stool specimens were examıned taken from patients with diarrhea whose ages ranged from 0 to 80 of applied to Hospital of Cumhuriyet University, Numune and Goverment Hospital in Sivas. All samples were tested for Cryptosporidium spp. G.intestinalis antigen by DFA and oocysts via gold Standard modified acidfast staining for Cryptosporidium spp.and G.intestinalis direct microscopy. Results. One specimen was found to be positive by modified acid-fast staining method and four specimens by DFA method were found to be positive for cryptosporidiosis. On the other hand two specimens were found to be positive both direct microscopy and DFA for giardiosis. Conclusion. The results of DFA test indicate that the simple, rapid, reliable, and standardized test is sensitive and specific for routine diagnosis and may be useful for large-scale epidemiological studies of cryptosporidiosis and giardiosis
References
- Ramirez NE Ward LA, Sreevatsan S. A review of the biology and epidemiology of cryptosporidiosis in humans and animals. Microbes Infect 2004; 6: 773-85.
- Tzipori S, Widmer G. A hundred year retrospective on cryptosporidiosis. Trends Parasitol 2008; 24: 184-9.
- Mosier DA, Oberst RD. Cryptosporidiosis. A global challenge. Ann NY Acad. Sci 2000; 916: 102-11.
- Richardson AJ, Frankenberg RA, Buck AC, Selcon JB, ColbourneJW, Mayon- White RT. An outbreak of waterborne cryptosporidiosis in Swindon and Oxfordshire. Epidemiol. Infect 1991; 107: 485-95.
- Mac Kenzie WR, Hoxie NJ, Proctor ME, Gradus MS, Blair KA, Peterson DE, Kazmierczak JJ, Addiss DG, Fox KR, Rose JB. A massive outbreak in Milwaukee of Cryptosporidium infection transmittedthrough the publicwater supply. N Eng J Med 1994; 331: 161-7.
- Çiçek M, Yılmaz H. İshalli çocuklarda Cryptosporidium spp. ve diğer barsak parazitlerinin yaygınlığı, Dicle Medical Journal 2011; 38: 70-5.
- Elgun G, Koltas IS. Investigation of Cryptosporidium spp.antigen by ELISA method in stool specimens obtained from patients with diarrhea. Parasitol.Res 2011; 108: 395-7.
- Yilmaz H, Tas Cengiz Z, Cicek M. Investigation of cryptosporidiosis by enzyme- linked immunosorbent assay and microscopy in children with diarrhea. Saudi Med J 2008; 29: 526-59.
- Ozcelik S, Poyraz Ö, Kalkan K, Malatyalı E, Degerli S. The investigation of Cryptosporidium spp. prevalence in cattle and farmers by ELISA. The Journal of the Faculty of Veterinary Medicine, University of Kafkas. SA 2012: 61-4.
- Özçelik S, Dökmetaş S, Sümer Z, İçağasıoğlu D, Dökmetaş İ. Gastroenteritlilerde Cryptosporidium görülme sıklığı Türk. Parazitol. Derg 1996; 20: 333-7.
- Çeliksöz A, Çelik S.Cumhuriyet Üniversitesi Hastanesi’nde gastroenteritli ve malnütrisyonlu hastalarda Cryptosporidium spp. Araştırması. Türk Parazitol Derg 2003; 27: 85-8.
- Graczyk TK, Kacprzak M, Neczaj E, Tamang L, Graczyk H, Lucy EF, Girouard AS. Occurrence of Cryptosporidium and Giardia in sewage sludge and solid waste landfill leachate and quantitative comparative analysis of sanitization treatments on pathogen inactivation. Environmental Research 2008; 106: 27-33.
- LeChevalier, MW, Norton, WD, Lee, RG. Occurrence of Giardia and Cryptosporidium spp. in surface water supplies. Applied and Environmental Microbiology 1991; 57: 2610-6.
- Lonigro A, Pollice A, Spinelli R, Berrilli F, Di Cave D, D’Orazi C, Cavallo P, Brandonisio O. Giardia cysts and Cryptosporidium oocysts in membrane-filtered municipal wastewater used for irrigation. Applıed and Envıronmental Mıcrobıology 2006; 72: 7916-78.
- Madore, MS, Rose, JB, Gerba, CP., Arrowood, MJ, & Sterling, CR. Occurrence of Cryptosporidium oocysts in sewage effluents and selected surface waters. Journal of Parasitology 1987; 73: 702-5.
- Üner A, Tanrıverdi S, Caner A, Değirmenci A. Cryptosporidium’larda Moleküler Biyolojik Yapı ve Çalışmalar. Moleküler Parazitoloji içinde. Ed.Özcel MA, Tanyüksel M, Eren H. Türkiye Parazitoloji Derneği Yayını, Meta Basım, İzmir 2009; 22.
- Ak M, Türk M, Güneş K. Giardiosis. Tıbbi Parazit Hastalıkları içinde. Ed: MA Özcel. Meta basım, İzmir 2007; 323-44.
- Uyar Y, Özkan AT. Amebiyazis, giardiyazis ve kriptosporidiyazis tanısında antijen tarama yöntemlerinin yeri. Türkiye Parazitoloji Derg 2009; 33: 140-50.
- Doğan N, Akgün Y. İshalli olgularda Cryptosporidium ookistlerinin araştırılması. Türk. Parazitol.Derg 1998; 22: 243-6.
- Aydın F, Katırcıoğlu İ, Köseahmet F, Bakır T, Bingöl R. Kronik diyareli hastaların dışkı örneklerinde Kriptosporidiumun belirlenmesi. İnfeksiyon Derg. 1995; 9: 151-5.
- Sönmez TG, Gülenç S. Diskida Cryptosporidium spp. antijenlerinin ELISA ile arastirilmasi. Türkiye Parazitol Derg 2008; 32: 198-201.
- Graczyk TK, Cranfield MR, Fayer R. Evaluation of commercial enzyme immunoassay (EIA) and immunofluorescent antibody (FA) test kits fot detection of Cryptosporidium oocysts of species other than Cryptosporidium parvum. Am J Trop Med Hyg 1996; 54: 274-9.
- Melrose W, Johnson K, Nimmo G. Use of multiplex real-time PCR to improve the detection of Giardia lamblia and Cryptosporidium parvum in human faecal samples. Amer.Soc. Trop. Med & Hygiene 2007: 77: 76.
- Özçelik S, Değerli S. Türkiye’de Giardiosis. T.Parazitol. Derg 1998; 22: 292-8.
- Muhsen K, Levine MM. A systematic review and meta-analysis of the association between Giardia lamblia and endemic pediatric diarrhea in developing countries. Clin.Infect Dis 2012; 55: 271-93.
- Veenemans J, Mank T, Ottenhof M, Baidjoe A, Mbugi E, Demir AY, Weilders JP, Savelkoul HF, Verhoef H. Protection against diarrhea associated with Giardia intestinalis is lost with multi-nutrient supplementation: A study in Tanzanian Children. PLoS Negl Trop Dis 2011; 5: 1158.
- Doğruman Al F, Kuştimur S, Özekinci T, Balaban N, İlhan MN. The use of Enzyme Linked Immunosorbent Assay (ELISA) and Direct Fluoresan Antibody (DFA) methods for diagnosis of Giardia intestinalis. Acta Parasitologica Turcica 2006; 30: 275-8.
- Weitzel T, Dittrch S, Möhl I, Adusu E, Jelinek T. Evaluation of seven commercial antigen detection tests for Giardia and Cryptosporidium in stool samples. Eur Soc Clin Micro İnf Dis 2006; 12: 656-9.
İshalli hastalarda direkt fluoresan antikor-dfa yöntemi ile giardia ve cryptosporidium spp. araştırılması
Abstract
Özet
Amaç. Cryptosporidium spp. ve Giardia intestinalis insanda ishal nedeni olabilen önemli protozoonlardır. Bu parazitozların tanısında etkensel tanı yöntemlerinin yanında antijen tarama testleri de kullanılabilmektedir. Çalışmamızda ishalli hastalarda konvansiyonel yöntemlerle birlikte Direkt Fluoresan Antikor (DFA) yöntemi ile G.intestinalis ve Cryptosporidium spp. varlığının saptanması ve bu iki protozoonun gastroenterit olgularındaki rolünün araştırılması amaçlanmıştır. Yöntem. Sivas il merkezindeki üç hastanenin (Cumhuriyet Üniversitesi Uygulama ve Araştırma Hastanesi, Sivas Numune ve Sivas Devlet Hastanesi) çeşitli servislerine enterit yakınmalarıyla başvuran 32’si kadın, 68’i erkek toplam 100 ishalli hastaya ait dışkı örneklerinde direkt inceleme ve asit-fast boyama yöntemi ile yapılan mikroskobik incelemelerde parazitlere ait kist ve/veya ookistler, DFA yöntemi ile parazitlere ait antijenler araştırılmıştır. İncelenen grupta yer alan hastalar 0-80 yaş aralığında bulunmaktadır. Bulgular. Çalışmada, ishalli 100 dışkı örneğinin 2’sinde direkt mikroskobide G.intestinalis görülmüştür. Asit-fast boyama ile birinde Cryptosporidium spp. görülürken, DFA ile 4’ünde (%4,0) Cryptosporidium spp., 2’sinde (%2,0) G. intestinalis antijenlerine rastlanmıştır. Cryptosporidium spp. saptanan hastaların erişkin yaş grubunda yer alması ilginç bulunmuştur. Sonuç. Cryptosporidium spp. ve G. intestinalis’in ikisini de içeren DFA testleri rutin çalışmalarda ve prevalans araştırmalarında kullanılabilen kolay, duyarlı ve güvenilir bir testtir. Diğer taraftan enterit nedenleri arasında sayılan bu iki protozoon incelenen hasta grubunda toplam %6 oranında bulunmuştur.
Anahtar sözcükler: Cryptosporidium spp., giardia intestinalis, DFA, ishal
Abstract
Aim. Cryptosporidium spp. and Giardia intestinalis are important parasitic protozoan causing diarrhea in developing and developed countries. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive and time-consuming. Thus, we aimed the presence of these protozoa in patients with diarrhea and to evaluate the usefulness of direct fluoresan antibody (DFA) test in detecting Cryptosporidium spp and G.intestinalis. from fecal specimens. Methods. For this aim, microscopy and specific antigen detection methods were used to determine Cryptosporidium spp. and G.intestinalis. One hundred stool specimens were examıned taken from patients with diarrhea whose ages ranged from 0 to 80 of applied to Hospital of Cumhuriyet University, Numune and Goverment Hospital in Sivas. All samples were tested for Cryptosporidium spp. G.intestinalis antigen by DFA and oocysts via gold Standard modified acid-fast staining for Cryptosporidium spp.and G.intestinalis direct microscopy. Results. One specimen was found to be positive by modified acid-fast staining method and four specimens by DFA method were found to be positive for cryptosporidiosis. On the other hand two specimens were found to be positive both direct microscopy and DFA for giardiosis. Conclusion. The results of DFA test indicate that the simple, rapid, reliable, and standardized test is sensitive and specific for routine diagnosis and may be useful for large-scale epidemiological studies of cryptosporidiosis and giardiosis.
Keywords: Cryptosporidium spp., giardia intestinalis, DFA, diarrhea
Keywords
References
- Ramirez NE Ward LA, Sreevatsan S. A review of the biology and epidemiology of cryptosporidiosis in humans and animals. Microbes Infect 2004; 6: 773-85.
- Tzipori S, Widmer G. A hundred year retrospective on cryptosporidiosis. Trends Parasitol 2008; 24: 184-9.
- Mosier DA, Oberst RD. Cryptosporidiosis. A global challenge. Ann NY Acad. Sci 2000; 916: 102-11.
- Richardson AJ, Frankenberg RA, Buck AC, Selcon JB, ColbourneJW, Mayon- White RT. An outbreak of waterborne cryptosporidiosis in Swindon and Oxfordshire. Epidemiol. Infect 1991; 107: 485-95.
- Mac Kenzie WR, Hoxie NJ, Proctor ME, Gradus MS, Blair KA, Peterson DE, Kazmierczak JJ, Addiss DG, Fox KR, Rose JB. A massive outbreak in Milwaukee of Cryptosporidium infection transmittedthrough the publicwater supply. N Eng J Med 1994; 331: 161-7.
- Çiçek M, Yılmaz H. İshalli çocuklarda Cryptosporidium spp. ve diğer barsak parazitlerinin yaygınlığı, Dicle Medical Journal 2011; 38: 70-5.
- Elgun G, Koltas IS. Investigation of Cryptosporidium spp.antigen by ELISA method in stool specimens obtained from patients with diarrhea. Parasitol.Res 2011; 108: 395-7.
- Yilmaz H, Tas Cengiz Z, Cicek M. Investigation of cryptosporidiosis by enzyme- linked immunosorbent assay and microscopy in children with diarrhea. Saudi Med J 2008; 29: 526-59.
- Ozcelik S, Poyraz Ö, Kalkan K, Malatyalı E, Degerli S. The investigation of Cryptosporidium spp. prevalence in cattle and farmers by ELISA. The Journal of the Faculty of Veterinary Medicine, University of Kafkas. SA 2012: 61-4.
- Özçelik S, Dökmetaş S, Sümer Z, İçağasıoğlu D, Dökmetaş İ. Gastroenteritlilerde Cryptosporidium görülme sıklığı Türk. Parazitol. Derg 1996; 20: 333-7.
- Çeliksöz A, Çelik S.Cumhuriyet Üniversitesi Hastanesi’nde gastroenteritli ve malnütrisyonlu hastalarda Cryptosporidium spp. Araştırması. Türk Parazitol Derg 2003; 27: 85-8.
- Graczyk TK, Kacprzak M, Neczaj E, Tamang L, Graczyk H, Lucy EF, Girouard AS. Occurrence of Cryptosporidium and Giardia in sewage sludge and solid waste landfill leachate and quantitative comparative analysis of sanitization treatments on pathogen inactivation. Environmental Research 2008; 106: 27-33.
- LeChevalier, MW, Norton, WD, Lee, RG. Occurrence of Giardia and Cryptosporidium spp. in surface water supplies. Applied and Environmental Microbiology 1991; 57: 2610-6.
- Lonigro A, Pollice A, Spinelli R, Berrilli F, Di Cave D, D’Orazi C, Cavallo P, Brandonisio O. Giardia cysts and Cryptosporidium oocysts in membrane-filtered municipal wastewater used for irrigation. Applıed and Envıronmental Mıcrobıology 2006; 72: 7916-78.
- Madore, MS, Rose, JB, Gerba, CP., Arrowood, MJ, & Sterling, CR. Occurrence of Cryptosporidium oocysts in sewage effluents and selected surface waters. Journal of Parasitology 1987; 73: 702-5.
- Üner A, Tanrıverdi S, Caner A, Değirmenci A. Cryptosporidium’larda Moleküler Biyolojik Yapı ve Çalışmalar. Moleküler Parazitoloji içinde. Ed.Özcel MA, Tanyüksel M, Eren H. Türkiye Parazitoloji Derneği Yayını, Meta Basım, İzmir 2009; 22.
- Ak M, Türk M, Güneş K. Giardiosis. Tıbbi Parazit Hastalıkları içinde. Ed: MA Özcel. Meta basım, İzmir 2007; 323-44.
- Uyar Y, Özkan AT. Amebiyazis, giardiyazis ve kriptosporidiyazis tanısında antijen tarama yöntemlerinin yeri. Türkiye Parazitoloji Derg 2009; 33: 140-50.
- Doğan N, Akgün Y. İshalli olgularda Cryptosporidium ookistlerinin araştırılması. Türk. Parazitol.Derg 1998; 22: 243-6.
- Aydın F, Katırcıoğlu İ, Köseahmet F, Bakır T, Bingöl R. Kronik diyareli hastaların dışkı örneklerinde Kriptosporidiumun belirlenmesi. İnfeksiyon Derg. 1995; 9: 151-5.
- Sönmez TG, Gülenç S. Diskida Cryptosporidium spp. antijenlerinin ELISA ile arastirilmasi. Türkiye Parazitol Derg 2008; 32: 198-201.
- Graczyk TK, Cranfield MR, Fayer R. Evaluation of commercial enzyme immunoassay (EIA) and immunofluorescent antibody (FA) test kits fot detection of Cryptosporidium oocysts of species other than Cryptosporidium parvum. Am J Trop Med Hyg 1996; 54: 274-9.
- Melrose W, Johnson K, Nimmo G. Use of multiplex real-time PCR to improve the detection of Giardia lamblia and Cryptosporidium parvum in human faecal samples. Amer.Soc. Trop. Med & Hygiene 2007: 77: 76.
- Özçelik S, Değerli S. Türkiye’de Giardiosis. T.Parazitol. Derg 1998; 22: 292-8.
- Muhsen K, Levine MM. A systematic review and meta-analysis of the association between Giardia lamblia and endemic pediatric diarrhea in developing countries. Clin.Infect Dis 2012; 55: 271-93.
- Veenemans J, Mank T, Ottenhof M, Baidjoe A, Mbugi E, Demir AY, Weilders JP, Savelkoul HF, Verhoef H. Protection against diarrhea associated with Giardia intestinalis is lost with multi-nutrient supplementation: A study in Tanzanian Children. PLoS Negl Trop Dis 2011; 5: 1158.
- Doğruman Al F, Kuştimur S, Özekinci T, Balaban N, İlhan MN. The use of Enzyme Linked Immunosorbent Assay (ELISA) and Direct Fluoresan Antibody (DFA) methods for diagnosis of Giardia intestinalis. Acta Parasitologica Turcica 2006; 30: 275-8.
- Weitzel T, Dittrch S, Möhl I, Adusu E, Jelinek T. Evaluation of seven commercial antigen detection tests for Giardia and Cryptosporidium in stool samples. Eur Soc Clin Micro İnf Dis 2006; 12: 656-9.
Details
| Primary Language | Turkish |
|---|---|
| Journal Section | Basic Science Research Articles |
| Authors | |
| Publication Date | October 14, 2014 |
| Published in Issue | Year 2014Volume: 36 Issue: 4 |