Ege Üniversitesi Tıp Fakültesi Hastanesinde Toksoplazmozis Sıklığının Real-Time PZR ile Belirlenmesi
Yıl 2022,
, 217 - 223, 30.06.2022
Muhammet Karakavuk
,
Hüseyin Can
,
Aysu Değirmenci Döşkaya
,
Adnan Gürüz
,
Mert Döşkaya
Öz
Toxoplasma gondii insan ve sıcakkanlı hayvanları enfekte edebilen zoonotik protozoon bir parazittir. Bu çalışmada çeşitli hasta gruplarından alınmış klinik örneklerde toksoplazmozis sıklığının araştırılması amaçlanmıştır. Çalışma kapsamında 2009-2019 yılları arasında Ege Üniversitesi Tıp Fakültesi Parazitoloji Anabilim Dalı Moleküler Parazitoloji laboratuvarına gönderilen ağırlıklı olarak amniyon sıvısı, kan ve beyin omurilik sıvısı örneklerinin dahil olduğu toplam 535 klinik örnek değerlendirilmiştir. Bu örneklerde DNA izolasyonu sonrası T. gondii RE geni varlığı Real-Time PZR ile araştırılmıştır.
Belirtilen zaman aralığında toksoplazmozis sıklığı %2,61 (14/535) olarak tespit edilmiştir. Hastalığın direkt
etkilediği merkezi organlardan alınan örneklerde pozitiflik oranı %5,40 (8/148) iken periferden alınan örneklerde pozitiflik oranı %1,74 (5/286) olarak tespit edilmiş ve bu fark istatistiksel olarak anlamlı bulunmuştur (P<0,05). İmmunsüpresif hastalarda ise hastalığın pozitiflik oranı daha yüksek olmasına rağmen istatistiksel olarak anlamlı bir fark saptanmamıştır (P>0,05).
Elde edilen bulgular toksoplazmozis tanısında Real-Time PZR yönteminin oldukça önemli bir yeri olduğunu
göstermektedir. Ayrıca, örnek alım yöntemlerinin hastalığın tanısında oldukça önemli olduğu sonucuna varılmıştır.
Destekleyen Kurum
Ege Üniversitesi
Proje Numarası
13-TIP-050
Teşekkür
Bu çalışma Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından 13-TIP-050 numaralı proje ile desteklenmiştir
Kaynakça
- Belaz, S., Gangneux, J. P., Dupretz, P., Guiguen, C., & Robert-Gangneuxa, F. (2015). A 10-year retrospective comparison of two target sequences, REP-529 and B1, for Toxoplasma gondii detection by quantitative PCR. Journal of Clinical Microbiology, 53(4), 1294–1300. https://doi.org/10.1128/JCM.02900-14
- Caner, A., Döşkaya, M., Karasu, Z., Değirmenci, A., Guy, E., Kılıç, M., … Gürüz, Y. (2007). Incidence and Diagnosis of Active Toxoplasma Infection Among Liver Transplant Recipients in Western Turkey. Liver Transplantation, 13(5), 767–768. https://doi.org/10.1002/lt
- Celik, T., Karaman, U., Celebi, B., Turan, A., Babur, C., & Daldal, N. (2008). Evaluation of the municipality dustmen in terms of toxoplasmosis and listeriosis seropozitivity in Malatya. Turk Hijyen ve Deneysel Biyoloji Dergisi, 65(2), 81–85.
- Costa, J. M., & Bretagne, S. (2012). Variation of B1 gene and AF146527 repeat element copy numbers according to Toxoplasma gondii strains assessed using real-time quantitative PCR. Journal of Clinical Microbiology, 50(4), 1452–1454. https://doi.org/10.1128/JCM.06514-11
- Döşkaya, M., Pullukçu, H., Karakavuk, M., Şahar, E. A., Taşbakan, M. S., Taşbakan, M. I., … Gürüz, A. Y. (2019). Comparison of an in house and a commercial real-time polymerase chain reaction targeting Toxoplasma gondii RE gene using various samples collected from patients in Turkey. BMC Infectious Diseases, 19(1). https://doi.org/10.1186/s12879-019-4666-z
- Döşkaya, Mert, Caner, A., Ajzenberg, D., Deǧirmenci, A., Dardé, M. L., Can, H., … Gürüz, Y. (2013). Isolation of Toxoplasma gondii strains similar to Africa 1 genotype in Turkey. Parasitology International, 62(5), 471–474. https://doi.org/10.1016/j.parint.2013.06.008
- Dubey, J. ., Miller, N. ., & J.K, F. (1970). The Toxoplasma gondii oocyst from cat feces. J Exp Med, 132(4), 636–662.
- Ertug, S., Okyay, P., Turkmen, M., & Yuksel, H. (2005). Seroprevalence and risk factors for toxoplasma infection among pregnant women in Aydin province, Turkey. BMC Public Health, 5, 1–6. https://doi.org/10.1186/1471-2458-5-66
- Filisetti, D., Sterkers, Y., Brenier-Pinchart, M. P., Cassaing, S., Dalle, F., Delhaes, L., … Bastien, P. (2015). Multicentric Comparative assessment of the bio-evolution Toxoplasma gondii detection kit with eight laboratory-developed PCR assays for molecular diagnosis of congenital toxoplasmosis. Journal of Clinical Microbiology, 53(1), 29–34. https://doi.org/10.1128/JCM.01913-14
- Gunel, T., Kalelioglu, I., Ermis, H., Has, R., & Aydinli, K. (2012). Large scale pre-diagnosis of Toxoplasma gondii DNA genotyping by real-time PCR on amniotic fluid. Biotechnology and Biotechnological Equipment, 26(2), 2913–2915. https://doi.org/10.5504/bbeq.2011.0106
- Hakko, E., Ozkan, H. A., Karaman, K., & Gulbas, Z. (2013). Analysis of cerebral toxoplasmosis in a series of 170 allogeneic hematopoietic stem cell transplant patients. Transplant Infectious Disease, 15(6), 575–580. https://doi.org/10.1111/tid.12138
- Harma, M., Harma, M., Güngen, N., & Demir, N. (2004). Toxoplasmosis in pregnant women in Şanlıurfa, Southeastern Anatolia City, Turkey. Journal of Egyptian Society for Parasitology, 34, 519–525.
- Holland, G. N. (2003). Ocular toxoplasmosis: A global reassessment. Part I: Epidemiology and course of disease. American Journal of Ophthalmology, 136(6), 973–988. https://doi.org/10.1016/j.ajo.2003.09.040
- Homan, W. ., Vercammen, M., Braekeleer, J. De, & Verschueren, H. (2000). Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. International Journal for Parasitology, 30(1), 69–75.
- Karakavuk, M., Aldemir, D., Mercier, A., Şahar, E. A., Can, H., Murat, J. B., … Döşkaya, M. (2018). Prevalence of toxoplasmosis and genetic characterization of Toxoplasma gondii strains isolated in wild birds of prey and their relation with previously isolated strains from Turkey. PLoS ONE, 13(4), 1–17. https://doi.org/10.1371/journal.pone.0196159
- Karakavuk, M., Aykur, M., Şahar, E. A., Karakuş, M., Aldemir, D., Döndüren, Ö., … Döşkaya, M. (2017). First time identification of Acanthamoeba genotypes in the cornea samples of wild birds; Is Acanthamoeba keratitis making the predatory birds a target? Experimental Parasitology, 183, 137–142. https://doi.org/10.1016/j.exppara.2017.08.007
- Karakavuk, M., Can, H., Karakavuk, T., Gül, A., Alak, S. E., Gül, C., … Döşkaya, A. D. (2022). Rapid detection of Toxoplasma gondii DNA in cat feces using colorimetric loop-mediated isothermal amplification (LAMP) assays targeting RE and B1 genes. Comparative Immunology, Microbiology and Infectious Diseases, 81(December 2021). https://doi.org/10.1016/j.cimid.2022.101745
- Karakavuk, M., Can, H., Selim, N., Yesilsiraz, B., Atli, E., Sahar, E. A., … Döskaya, M. (2021). Investigation of the role of stray cats for transmission of toxoplasmosis to humans and animals living in Izmir, Turkey. Journal of Infection in Developing Countries, 15(1), 155–162. https://doi.org/10.3855/jidc.13932
- Kim, K., & Weiss, L. M. (2004). Toxoplasma gondii: The model apicomplexan. International Journal for Parasitology, 34(3), 423–432. https://doi.org/10.1016/j.ijpara.2003.12.009
- Kompalic-Cristo, A., Frotta, C., Suárez-Mutis, M., Fernandes, O., & Britto, C. (2007). Evaluation of a real-time PCR assay based on the repetitive B1 gene for the detection of Toxoplasma gondii in human peripheral blood. Parasitology Research, 101(3), 619–625. https://doi.org/10.1007/s00436-007-0524-9
- Maani, S., Rezanezhad, H., Solhjoo, K., Kalantari, M., & Erfanian, S. (2021). Genetic characterization of Toxoplasma gondii isolates from human spontaneous aborted fetuses in Jahrom, southern Iran. Microbial Pathogenesis, 161(PA), 105217. https://doi.org/10.1016/j.micpath.2021.105217
- Montoya, J., & Liesenfeld, O. (2004). Toxoplasmosis. Lancet, 363, 1965–1976. https://doi.org/10.4067/s0718-04622007000200009
- Remington, J. S., Thulliez, P., & Montoya, J. G. (2004). Recent Developments for Diagnosis of Toxoplasmosis. Journal of Clinical Microbiology, 42(3), 941–945. https://doi.org/10.1128/JCM.42.3.941-945.2004
- Robert-Gangneux, F., & Dardé, M. L. (2012). Epidemiology of and diagnostic strategies for toxoplasmosis. Clinical Microbiology Reviews, 25(2), 264–296. https://doi.org/10.1128/CMR.05013-11
- Salant, H., Spira, D. T., & Hamburger, J. (2010). A comparative analysis of coprologic diagnostic methods for detection of toxoplama gondii in cats. American Journal of Tropical Medicine and Hygiene, 82(5), 865–870. https://doi.org/10.4269/ajtmh.2010.09-0635
- Sert, U. Y., Ozgu-Erdinc, A. S., Gokay, S., & Engin-Ustun, Y. (2019). Toxoplasma Screening Results of 84587 Pregnant Women in a Tertiary Referral Center in Turkey. Fetal and Pediatric Pathology, 38(4), 307–316. https://doi.org/10.1080/15513815.2019.1587122
- Tekay, F., & Özbek, E. (2007). Çi ğ Köftenin Yayg ı n Tüketildi ğ i Ş anl ı urfa İ linde Kad ı nlarda Toxoplasma gondii Seroprevalans ı. 31(3), 176–179.
- Wahab, T., Edvinsson, B., Palm, D., & Lindh, J. (2010). Comparison of the AF146527 and B1 repeated elements, two real-time PCR targets used for detection of Toxoplasma gondii. Journal of Clinical Microbiology, 48(2), 591–592. https://doi.org/10.1128/JCM.01113-09
Determination of Toxoplasmosis Frequency by Real-Time PCR in Ege University Medical Faculty Hospital
Yıl 2022,
, 217 - 223, 30.06.2022
Muhammet Karakavuk
,
Hüseyin Can
,
Aysu Değirmenci Döşkaya
,
Adnan Gürüz
,
Mert Döşkaya
Öz
Toxoplasma gondii is a zoonotic protozoan parasite that can infect humans and warm-blooded animals. In this study, it was aimed to investigate the frequency of toxoplasmosis in clinical samples taken from various patient groups. Within the scope of the study, a total of 535 clinical samples, including mainly amniotic fluid, blood and cerebrospinal fluid samples, which were sent to the Molecular Parasitology laboratory of Ege University Faculty of Medicine, Department of Parasitology between 2009 and 2019, were evaluated.
The frequency of toxoplasmosis was determined as 2.61% (14/535) in the specified time period. While the positivity rate was 5.40% (8/148) in the samples taken from the central organs directly affected by the disease, the positivity rate was 1.74% (5/286) in the samples taken from the periphery, and this difference was statistically significant (P<0.05). Although the positivity rate of the disease was higher in immunosuppressive patients, no statistically significant difference was found (P>0.05).
The findings show that Real-Time PCR method has a very important place in the diagnosis of active toxoplasmosis. In addition, it was concluded that sampling methods are very important in the diagnosis of the disease.
Proje Numarası
13-TIP-050
Kaynakça
- Belaz, S., Gangneux, J. P., Dupretz, P., Guiguen, C., & Robert-Gangneuxa, F. (2015). A 10-year retrospective comparison of two target sequences, REP-529 and B1, for Toxoplasma gondii detection by quantitative PCR. Journal of Clinical Microbiology, 53(4), 1294–1300. https://doi.org/10.1128/JCM.02900-14
- Caner, A., Döşkaya, M., Karasu, Z., Değirmenci, A., Guy, E., Kılıç, M., … Gürüz, Y. (2007). Incidence and Diagnosis of Active Toxoplasma Infection Among Liver Transplant Recipients in Western Turkey. Liver Transplantation, 13(5), 767–768. https://doi.org/10.1002/lt
- Celik, T., Karaman, U., Celebi, B., Turan, A., Babur, C., & Daldal, N. (2008). Evaluation of the municipality dustmen in terms of toxoplasmosis and listeriosis seropozitivity in Malatya. Turk Hijyen ve Deneysel Biyoloji Dergisi, 65(2), 81–85.
- Costa, J. M., & Bretagne, S. (2012). Variation of B1 gene and AF146527 repeat element copy numbers according to Toxoplasma gondii strains assessed using real-time quantitative PCR. Journal of Clinical Microbiology, 50(4), 1452–1454. https://doi.org/10.1128/JCM.06514-11
- Döşkaya, M., Pullukçu, H., Karakavuk, M., Şahar, E. A., Taşbakan, M. S., Taşbakan, M. I., … Gürüz, A. Y. (2019). Comparison of an in house and a commercial real-time polymerase chain reaction targeting Toxoplasma gondii RE gene using various samples collected from patients in Turkey. BMC Infectious Diseases, 19(1). https://doi.org/10.1186/s12879-019-4666-z
- Döşkaya, Mert, Caner, A., Ajzenberg, D., Deǧirmenci, A., Dardé, M. L., Can, H., … Gürüz, Y. (2013). Isolation of Toxoplasma gondii strains similar to Africa 1 genotype in Turkey. Parasitology International, 62(5), 471–474. https://doi.org/10.1016/j.parint.2013.06.008
- Dubey, J. ., Miller, N. ., & J.K, F. (1970). The Toxoplasma gondii oocyst from cat feces. J Exp Med, 132(4), 636–662.
- Ertug, S., Okyay, P., Turkmen, M., & Yuksel, H. (2005). Seroprevalence and risk factors for toxoplasma infection among pregnant women in Aydin province, Turkey. BMC Public Health, 5, 1–6. https://doi.org/10.1186/1471-2458-5-66
- Filisetti, D., Sterkers, Y., Brenier-Pinchart, M. P., Cassaing, S., Dalle, F., Delhaes, L., … Bastien, P. (2015). Multicentric Comparative assessment of the bio-evolution Toxoplasma gondii detection kit with eight laboratory-developed PCR assays for molecular diagnosis of congenital toxoplasmosis. Journal of Clinical Microbiology, 53(1), 29–34. https://doi.org/10.1128/JCM.01913-14
- Gunel, T., Kalelioglu, I., Ermis, H., Has, R., & Aydinli, K. (2012). Large scale pre-diagnosis of Toxoplasma gondii DNA genotyping by real-time PCR on amniotic fluid. Biotechnology and Biotechnological Equipment, 26(2), 2913–2915. https://doi.org/10.5504/bbeq.2011.0106
- Hakko, E., Ozkan, H. A., Karaman, K., & Gulbas, Z. (2013). Analysis of cerebral toxoplasmosis in a series of 170 allogeneic hematopoietic stem cell transplant patients. Transplant Infectious Disease, 15(6), 575–580. https://doi.org/10.1111/tid.12138
- Harma, M., Harma, M., Güngen, N., & Demir, N. (2004). Toxoplasmosis in pregnant women in Şanlıurfa, Southeastern Anatolia City, Turkey. Journal of Egyptian Society for Parasitology, 34, 519–525.
- Holland, G. N. (2003). Ocular toxoplasmosis: A global reassessment. Part I: Epidemiology and course of disease. American Journal of Ophthalmology, 136(6), 973–988. https://doi.org/10.1016/j.ajo.2003.09.040
- Homan, W. ., Vercammen, M., Braekeleer, J. De, & Verschueren, H. (2000). Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. International Journal for Parasitology, 30(1), 69–75.
- Karakavuk, M., Aldemir, D., Mercier, A., Şahar, E. A., Can, H., Murat, J. B., … Döşkaya, M. (2018). Prevalence of toxoplasmosis and genetic characterization of Toxoplasma gondii strains isolated in wild birds of prey and their relation with previously isolated strains from Turkey. PLoS ONE, 13(4), 1–17. https://doi.org/10.1371/journal.pone.0196159
- Karakavuk, M., Aykur, M., Şahar, E. A., Karakuş, M., Aldemir, D., Döndüren, Ö., … Döşkaya, M. (2017). First time identification of Acanthamoeba genotypes in the cornea samples of wild birds; Is Acanthamoeba keratitis making the predatory birds a target? Experimental Parasitology, 183, 137–142. https://doi.org/10.1016/j.exppara.2017.08.007
- Karakavuk, M., Can, H., Karakavuk, T., Gül, A., Alak, S. E., Gül, C., … Döşkaya, A. D. (2022). Rapid detection of Toxoplasma gondii DNA in cat feces using colorimetric loop-mediated isothermal amplification (LAMP) assays targeting RE and B1 genes. Comparative Immunology, Microbiology and Infectious Diseases, 81(December 2021). https://doi.org/10.1016/j.cimid.2022.101745
- Karakavuk, M., Can, H., Selim, N., Yesilsiraz, B., Atli, E., Sahar, E. A., … Döskaya, M. (2021). Investigation of the role of stray cats for transmission of toxoplasmosis to humans and animals living in Izmir, Turkey. Journal of Infection in Developing Countries, 15(1), 155–162. https://doi.org/10.3855/jidc.13932
- Kim, K., & Weiss, L. M. (2004). Toxoplasma gondii: The model apicomplexan. International Journal for Parasitology, 34(3), 423–432. https://doi.org/10.1016/j.ijpara.2003.12.009
- Kompalic-Cristo, A., Frotta, C., Suárez-Mutis, M., Fernandes, O., & Britto, C. (2007). Evaluation of a real-time PCR assay based on the repetitive B1 gene for the detection of Toxoplasma gondii in human peripheral blood. Parasitology Research, 101(3), 619–625. https://doi.org/10.1007/s00436-007-0524-9
- Maani, S., Rezanezhad, H., Solhjoo, K., Kalantari, M., & Erfanian, S. (2021). Genetic characterization of Toxoplasma gondii isolates from human spontaneous aborted fetuses in Jahrom, southern Iran. Microbial Pathogenesis, 161(PA), 105217. https://doi.org/10.1016/j.micpath.2021.105217
- Montoya, J., & Liesenfeld, O. (2004). Toxoplasmosis. Lancet, 363, 1965–1976. https://doi.org/10.4067/s0718-04622007000200009
- Remington, J. S., Thulliez, P., & Montoya, J. G. (2004). Recent Developments for Diagnosis of Toxoplasmosis. Journal of Clinical Microbiology, 42(3), 941–945. https://doi.org/10.1128/JCM.42.3.941-945.2004
- Robert-Gangneux, F., & Dardé, M. L. (2012). Epidemiology of and diagnostic strategies for toxoplasmosis. Clinical Microbiology Reviews, 25(2), 264–296. https://doi.org/10.1128/CMR.05013-11
- Salant, H., Spira, D. T., & Hamburger, J. (2010). A comparative analysis of coprologic diagnostic methods for detection of toxoplama gondii in cats. American Journal of Tropical Medicine and Hygiene, 82(5), 865–870. https://doi.org/10.4269/ajtmh.2010.09-0635
- Sert, U. Y., Ozgu-Erdinc, A. S., Gokay, S., & Engin-Ustun, Y. (2019). Toxoplasma Screening Results of 84587 Pregnant Women in a Tertiary Referral Center in Turkey. Fetal and Pediatric Pathology, 38(4), 307–316. https://doi.org/10.1080/15513815.2019.1587122
- Tekay, F., & Özbek, E. (2007). Çi ğ Köftenin Yayg ı n Tüketildi ğ i Ş anl ı urfa İ linde Kad ı nlarda Toxoplasma gondii Seroprevalans ı. 31(3), 176–179.
- Wahab, T., Edvinsson, B., Palm, D., & Lindh, J. (2010). Comparison of the AF146527 and B1 repeated elements, two real-time PCR targets used for detection of Toxoplasma gondii. Journal of Clinical Microbiology, 48(2), 591–592. https://doi.org/10.1128/JCM.01113-09