Brusellozis tanısında polimeraz zincir reaksiyonunun etkinliğinin serolojik yöntemlerle karşılaştırılması ve virülans genlerinin saptanması
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Dosyalar
Tarih
2019
Yazarlar
Dergi Başlığı
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Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Bruselloz, çoklu organ tutulumu ile ilerleyen zoonotik bir enfeksiyondur. Klasik tanıda kullanılan kültür ve serolojik testler uzun süre gerektirmekte ve yanlışnegatif olarak sonuçlanabilmektedir. Bu çalışmada, hızla gelişen ve birkaç saatte sonuçlanabilen bir tanı yöntemi olan PCR'ın bruselloz tanısındaki etkinliği araştırılmıştır. Materyal ve Metot: Çalışmaya, hastanemizde 01 Ocak 2018 ile 31 Temmuz 2018 tarihleri arasında saptanan 60 hasta ve 60 kontrol numunesi dahil edildi. Numunelerde real-time PCR yöntemi ile Brucella spp. DNA'sı araştırıldı ve pozitif örneklerde PCR yöntemi ile tür ayrımı ve virülans genleri çalışıldı. PCR yönteminin tanı etkinlik değerleri hesaplandı. Bulgular: Hasta numunelerinin sekizinin kan kültüründe Brucella spp. üremesi oldu. Serolojik yöntemlerle antikor saptanan 60 hastanın 50'sinde akut hastalık göstergesi olan ≥1/160 titre değeri mevcuttu ve bu hastaların 32'si akut hasta, geri kalan 18'i ise tedavi altındaki hastalardı. Yapılan real-time PCR testi ile hasta grubunda 19 örnekte Brucella spp. DNA'sı saptanırken, bu türlerin hepsi Brucella melitensis olarak belirlendi Pozitif örneklerin hepsinde omp19, wbkA, manA, mviN, ure ve perA virülans genleri pozitif bulundu. PCR'ın hasta grubu ve akut hastalar için saptanan duyarlılığı sırasıyla %31.6 ve %59.3; özgüllük ve Pozitif Prediktif Değeri %100 ve akut hastalar için Negatif Prediktif Değeri ise %87.1 olarak hesaplandı. Sonuç: Çalışmamızda, Brucella PCR testi ile akut hastaların yaklaşık %60'ının tanısının yapılabildiği, bu testin özgüllüğünün mükemmel düzeyde olduğu, olgularının hepsine B. melitensis'in neden olduğu ve suşların tüm önemli virülans genlerini taşıdığı saptanmıştır. Daha hassas saptama limiti olan PCR kitlerinin geliştirilmesi ve hastadan uygun dönemde numune alınması ile PCR'ın bruselloz tanısındaki etkinliği artacaktır. Anahtar Kelimeler: Brucella spp., Bruselloz, Serolojik Testler, PCR, Virülans.
Aim: Brucellosis is a zoonotic infection that progress with multiorgan involvement. The classical diagnostic methods, including culture and serological tests, require long time to be completed and may yield false negative results. In this study, the performance of PCR, as a rapidly-developing method that is able to give results within hours, was aimed to be evaluated on the diagnosis of brucellosis. Material and Method: A total 60 patients and 60 control patients' samples which were assessed between January 1st, 2018 and July 31th, 2018 in our hospital were included. Brucella spp. DNA was investigated in the clinical samples with real-time PCR method, and species typing and virulence genes were studied with PCR. The diagnostic efficacy of PCR method was analyzed. Results: Brucella spp. growth occurred in eight of the patients' blood cultures. Fifty out of 60 patient samples which were found positive for antibody were determined as having ≥1/160 serum titer, and 32 of them having acute disease. Resting 18 patients were accepted as the patients who were currently under treatment. In the real-time PCR analysis, DNA of Brucella spp. were found positive in 19 samples in the patient group. Brucella melitensis were identified in all PCR-positive samples, and all these samples were also found positive for all the studied virulence genes including omp19, wbkA, manA, mviN, ure and perA. Of the PCR method, the sensitivity for patients group and acute patients were found as 31.6% and 59.3%, respectively; specificity and Positive Predictive values were determined as 100%, and Negative Predictive Value for acute patients was found as 87.1%. Conclusion: In this study, it was determined that Brucella PCR could diagnose almost 60% of acute patients, the specificity of this test was excellent, B. melitensis was the causative agent in all patients and all these strains were carrying all substantial virulence genes. It is concluded that more sensitive PCR kits having lower detection limit are required, and sampling from the patients in appropriate time will increase the efficacy of PCR in the diagnosis of brucellosis. Key Words: Brucella spp., Brucellosis, Serological Tests, PCR, Virulence.
Aim: Brucellosis is a zoonotic infection that progress with multiorgan involvement. The classical diagnostic methods, including culture and serological tests, require long time to be completed and may yield false negative results. In this study, the performance of PCR, as a rapidly-developing method that is able to give results within hours, was aimed to be evaluated on the diagnosis of brucellosis. Material and Method: A total 60 patients and 60 control patients' samples which were assessed between January 1st, 2018 and July 31th, 2018 in our hospital were included. Brucella spp. DNA was investigated in the clinical samples with real-time PCR method, and species typing and virulence genes were studied with PCR. The diagnostic efficacy of PCR method was analyzed. Results: Brucella spp. growth occurred in eight of the patients' blood cultures. Fifty out of 60 patient samples which were found positive for antibody were determined as having ≥1/160 serum titer, and 32 of them having acute disease. Resting 18 patients were accepted as the patients who were currently under treatment. In the real-time PCR analysis, DNA of Brucella spp. were found positive in 19 samples in the patient group. Brucella melitensis were identified in all PCR-positive samples, and all these samples were also found positive for all the studied virulence genes including omp19, wbkA, manA, mviN, ure and perA. Of the PCR method, the sensitivity for patients group and acute patients were found as 31.6% and 59.3%, respectively; specificity and Positive Predictive values were determined as 100%, and Negative Predictive Value for acute patients was found as 87.1%. Conclusion: In this study, it was determined that Brucella PCR could diagnose almost 60% of acute patients, the specificity of this test was excellent, B. melitensis was the causative agent in all patients and all these strains were carrying all substantial virulence genes. It is concluded that more sensitive PCR kits having lower detection limit are required, and sampling from the patients in appropriate time will increase the efficacy of PCR in the diagnosis of brucellosis. Key Words: Brucella spp., Brucellosis, Serological Tests, PCR, Virulence.
Açıklama
Anahtar Kelimeler
Mikrobiyoloji, Microbiology
Kaynak
WoS Q Değeri
Scopus Q Değeri
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Künye
Seyhan, Engin Vedat (2019). Brusellozis tanısında polimeraz zincir reaksiyonunun etkinliğinin serolojik yöntemlerle karşılaştırılması ve virülans genlerinin saptanması. Yayımlanmış Yüksek Lisans tezi, İnönü Üniversitesi, Malatya.1-77 ss.
