İndirekt Hemaglütinasyon Yöntemiyle anti-Echinococcus granulosus Antikorları Pozitif Saptanan Serum Örneklerinin Western Blot Testi ile Değerlendirilmesi

Öz

Amaç: Echinococcus granulosus larvasının insanlarda sebep olduğu kistik ekinokokkoz KE , ülkemizde hayvancılığın yoğun olarak yapıldığı bölgelerde yaygın görülen ve tanı için klinisyen, radyolog ve mikrobiyologların multidisipliner yaklaşımını gerektiren bir zoonozdur. Enzim-linked immünassay ELISA ve indirekt hemaglütinasyon IHA , hastaların tanısında ilk sırada tercih edilirken Western Blot WB testi daha çok doğrulama amacıyla kullanılmaktadır. Fakat serolojik testlerin hastalığın tanı ve takibinde tek başına kullanımı, değişken duyarlılık ve özgüllük oranları nedeniyle önerilmemekte, uygun tanı için birden fazla serolojik testin kullanımı gerekmektedir. Bu çalışmada, Aralık 2015-2016 tarihleri arasında Gazi Üniversitesi Tıp Fakültesi Mikrobiyoloji Laboratuvarı’na KE şüphesiyle gönderilen hasta serumlarında IHA yöntemiyle titrasyon veren örneklerin doğrulama testi olarak kabul edilen WB yöntemi ile değerlendirilmesi ve iki test arasındaki tutarlılığın saptanması amaçlanmıştır.Yöntem: KE şüpheli örnekler, E. granulosus antijenleri ile hazırlanmış IHA Fumouze Laboratoires

Anahtar Kelimeler

• Kistik ekinokokoz, seroloji, tanı, indirekt hemaglütinasyon testi, western blot

Western blot assay of anti-Echinococcus granulosus antibody positive serum samples by indirect haemagglutination method

Öz

Objective: Cystic echinococcosis CE is a zoonotic disease mainly caused by the larvae of Echinococcus granulosus which is common in rural areas in Turkey. A multidisciplinary approach consisting of clinicians, radiologists and microbiologists is required for the proper diagnosis of the disease. Enzyme-linked immunosorbent assay ELISA and indirect hemagglutination IHA tests are preferred in the primer diagnosis of cystic echinococcosis CE , while western blot WB is used to confirm the disease. However the use of serologic tests alone in diagnosis and follow-up of the disease is not recommended due to variable sensitivity and specificity rates and multiple serologic tests are required for appropriate diagnosis. In this study, it was aimed to compare the test results of patients sera sent to Gazi University Medical Faculty Microbiology Laboratory, between December 2015 and December 2016, with the preliminary diagnosis of CE, by WB test after those titrated with IHA. It is also aimed to determine the consistency between the two tests.Methods: CE suspicious specimens were first tested

Anahtar Kelimeler

• Cystic echinococcosis, serology, diagnosis, indirect haemagglutination test, western blot

Kaynakça

1. 1. Saeed I, Kapel C, Saida LA, Willingham L, Nansen P. Epidemiology of Echinococcus granulosus in Arbil province, northern Iraq, 1990-1998. J Helminthol, 2000; 74 (1): 83-88.
2. 2. Yazar S, Ozkan AT, Hökelek M, Polat E, Yilmaz H et al. Cystic echinococcosis in Turkey from 2001- 2005. Turkiye Parazitol Derg, 2008; 32 (3): 208-220.
3. 3. Açikgöz D, Inceboz T, Ozkara E, Korkmaz M, Birgen N, Uzün I. The investigation of frequency of cystic echinococcosis in the autopsies committed in the Speciality Department of Istanbul Forensic Medicine Institute. Turkiye Parazitol Derg, 2009;33(2):155- 157.
4. 4. Feng X, Qi X, Yang L, Duan X, Fang B et al. Human cystic and alveolar echinococcosis in the Tibet Autonomous Region (TAR), China. J Helminthol, 2015; 89 (6): 671-679. doi: 10.1017/ S0022149X15000656.
5. 5. Ito A, Budke CM. The present situation of echinococcoses in Mongolia. J Helminthol, 2015; 89 (6): 680-688. doi: 10.1017/S0022149X15000620.
6. 6. Raimkylov KM, Kuttubaev OT, Toigombaeva VS. Epidemiological analysis of the distribution of cystic and alveolar echinococcosis in Osh Oblast in the Kyrgyz Republic, 2000-2013. J Helminthol, 2015; 89 (6): 651-654. doi: 10.1017/S0022149X15000565.
7. 7. Agudelo-Higuita NI, Brunetti E, McCloskey C. Cystic echinococcosis. J Clin Microbiol, 2016; 54: 518-523. doi: 10.1128/JCM.02420-15.
8. 8. Deplazes P, Rinaldi L, Alvarez-Rojas CA, Torgerson PR, Harandi MF et al. Global distribution of alveolar and cystic echinococcosis. Adv Parasitol, 2017; 95: 315-493. doi: 10.1016/bs.apar.2016.11.001.

9. 9. Yaman İ, İnceboz Ü, İnceboz T, Keyik B, Uzgören E. Primary pelvic cystic echinococcosis, Turkiye Parazitol Derg. 2015;39(2):167-70. doi:10.5152/ tpd.2015.3623.
10. 10. Arda B, Pullukçu H, Yamazhan T, Sipahi OR, Tamsel S et al. Prevalence of Echinococcus granulosus detected using enzyme immunoassay and abdominal ultrasonography in a group of students staying in a state dormitory in Turkey. Turk J Med Sci, 2009; 39 (5): 791-794. doi:10.3906/sag-0805-76.
11. 11. Canda MŞ, Güray M, Canda T, Astarcioğlu H. The pathology of echinococcosis and the current echinococcosis problem in western Turkey (A report of pathologic features in 80 cases). Turk J Med Sci, 2003; 33 (6) 369-74.
12. 12. Eckert J, Gemmell MA, Meslin FX, Pawłowski, ZS. WHO Manual on echinococcosis in humans and animals: A public health problem of global concern 2002: 265 pp.
13. 13. Del Carpio M, Mercapide CH, Salvitti JC, Uchiumi L, Sustercic J et al. Early diagnosis, treatment and follow-up of cystic echinococcosis in remote rural areas in Patagonia: impact of ultrasound training of non-specialists. PLoS Negl Trop Dis, 2012; 6: e1444. doi: 10.1371/journal.pntd.0001444.
14. 14. Moro P, Schantz PM. Echinococcosis: a review. Int J Infect Dis, 2009; 13: 125-133. doi: 10.1016/j. ijid.2008.03.037.
15. 15. Biava MF, Dao A, Fortier B. Laboratory diagnosis of cystic hydatic disease. World J Surg, 2001; 25: 10-14.
16. 16. Güreser AS, Ozcan O, Ozunel L, Boyacioglu Zİ, Taylan-Ozkan, A. Evaluation of the radiological, biochemical and serological parameters of patients pre-diagnosed as cystic echinococcosis in Çorum, Turkey. Mikrobiyol Bul, 2015; 49(2): 231-239.
17. 17. Cherradi Y, Afifi R, Khannoussi W, Firwana M, Rahaoui A, Benazzouz M et al. Long-term results of percutaneous management of liver hydatid cysts: Experience of university hospital in endemic region. J Med Sur Res, 2016; 3 (2): 275-81.
18. 18. Siles-Lucas M, Casulli A, Conraths FJ, Müller N. Laboratory diagnosis of Echinococcus spp. in human patients and infected animals. Adv Parasitol, 2017; 96: 159-257. doi: 10.1016/bs.apar.2016.09.003.
19. 19. Yılmaz GR, Babür C. Diagnosis of echinococcosis. Turk Hij Den Biyol Derg, 2007; 64: 35-44.
20. 20. Kılıç S, Babur C, Taylan-Ozkan A. Comparison of the results of indirect hemagglutination and ELISA methods for the cases prediagnosed as hydatid cyst disease. Mikrobiyol Bul, 2007; 41: 571-577.
21. 21. Jin Y, Anvarov K, Khajibaev A, Hong S, Hong ST. Serodiagnosis of echinococcosis by ELISA using cystic fluid from Uzbekistan sheep. Korean J Parasitol, 2013; 51(3): 313-317. doi: 10.3347/ kjp.2013.51.3.313.
22. 22. Beyhan YE, Babür C, Mungan M, Özkan-Taylan A. Evaluation of cystic echinococcosis suspected patients applied to National Parasitology Reference Laboratory of Public Health Institution of Turkey between 2009-2013. Turkiye Parazitol Derg, 2015; 39(1): 17-21. doi: 10.5152/tpd.2015.3646.
23. 23. Chirag S, Fomda BA, Khan A, Malik AA, Lone GN et al. Detection of hydatid-specific antibodies in the serum and urine for the diagnosis of cystic echinococcosis in patients from the Kashmir Valley, India. J Helminthol, 2015; 89 (2): 232-237. doi: 10.1017/S0022149X13000837.
24. 24. Doiz O, Benito R, Younes S, Osuna A, Clavel A, Gomez-Lusa R. Western blot applied to the diagnosis and post-treatment monitoring of human hydatidosis. Diagn Microbiol Infect Dis, 2001; 41: 139-42.
25. 25. Aslan M, Yüksel P, Polat E, Cakan H, Ergin S et al. The diagnostic value of Western blot method in patients with cystic echinococcosis. New Microbiol, 2011; 34: 173-177.
26. 26. Reiter-Owona I, Grüner B, Frosch M, Hoerauf A, Kern P, Tappe D. Serological confirmatory testing of alveolar and cystic echinococcosis in clinical practice: results of a comparative study with commercialized and in-house assays. Clin Lab, 2009; 55 (1-2): 41-8.
27. 27. Manzano R, Sánchez-Ovejero C, HernándezGonzález A, Casulli A, Siles-Lucas, M. Serological diagnosis and follow-up of human cystic echinococcosis: a new hope for the future? Biomed Res Int, 2015;1-9. doi: 10.1155/2015/428205.
28. 28. Gwet KL. Handbook of Inter-Rate Reliability. 4th ed. Gaithersburg, USA. 2014.
29. 29. Kern P, Menezes da Silva A, Akhan O, Müllhaupt B, Vizcaychipi KA et al. The Echinococcoses: Diagnosis, clinical management and burden of disease. Adv Parasitol, 2017; 96: 259-369. doi: 10.1016/ bs.apar.2016.09.006.
30. 30. Zhang W, Li J, Mc Manus DP. M. Concepts in immunology and diagnosis of hydatid disease. Clin Microbiol Rev, 2003 16(1):18-36.
31. 31. Craig PS, Mc Manus DP, Lightowlers MW, Chabalgoity JA, Garcia HH et al. Prevention and control of cystic echinococcosis. Lancet Infect Dis, 2007; 7(6): 385- 394. doi: 10.1016/S1473-3099(07)70134-2.
32. 32. Carmena D, Benito A, Eraso E. Antigens for the immunodiagnosis of Echinococcus granulosus infection: An update. Acta Trop, 2006: 98; 74-86. doi: 10.1016/j.actatropica.2006.02.002.
33. 33. Auer H, Stöckl C, Suhendra S, Schneider R. Sensitivity and specificity of new commercial tests for the detection of specific Echinococcus antibodies. Wien Klin Wochenschr, 2009; 121: 37- 41. doi: 10.1007/s00508-009-1233-4.
34. 34. Hernández-González A, Santivañez S, García HH, Rodríguez S, Muñoz S et al. Improved serodiagnosis of cystic echinococcosis using the new recombinant 2B2t antigen. PLoS Negl Trop Dis, 2012; 6(7): e1714. doi: 10.1371/journal.pntd.0001714.
35. 35. Logar J, Soba B, Kotar T. Serological evidence for human cysticechinococcosis in Slovenia. BMC Infect Dis, 2008; 8: 4-7. doi: 10.1186/1471-2334-8-63.
36. 36. Cetinkaya U, Hamamcı B, Kaya M, Gücüyetmez S, Kuk S et al. Investigation of anti-Echinococcus granulosus antibodies in patients with suspected cystic echinococcosis. Turkiye Parazitol Derg, 2012; 36(2): 57-60.
37. 37. Kanwar JR, Kanwar RK, Grewal AS, Vinayak, VK. Significance of detection of immune-complexed 8 kDa hydatid-specific antigen for immunodiagnosis of hydatidosis. FEMS Immunol Med Microbiol, 1994; 9: 231-236.
38. 38. Sarkari B, Rezaei Z. Immunodiagnosis of human hydatid disease: Where do we stand? World J Methodol, 2015; 5: 185-195. doi: 10.5662/wjm. v5.i4.185.
39. 39. Akcam AT, Ulku A, Koltas IS, Izol V, Bicer OS et al. Clinical characterisation of unusual cystic echinococcosis in Southern part of Turkey. Ann Saudi Med, 2014; 34: 508-516. doi: 10.5144/0256-4947.2014.508.
40. 40. Wuestenberg J, Gruener B, Oeztuerk S, Mason RA, Haenle MM et al. Diagnostics in cystic echinococcosis: serology versus ultrasonography. Turk J Gastroenterol, 2014; 25(4): 398-404. doi: 10.5152/tjg.2014.7112