GASTROİNTESTİNAL ŞİKÂYETİ OLAN HASTALARDA BAĞIRSAK PATOJENİ PROTOZOONLARIN PREVALANSININ MİKROSKOPİ VE MULTİPLEKS REAL-TIME PCR İLE ARAŞTIRILMASI

Yıl 2024, Cilt: 7 Sayı: 3, 180 - 184, 30.10.2024

Burak Karacan , Özden Boral , Gülay İmadoğlu Yetkin , Asli Cifcibasi Ormeci , Ozan Özkaya , Murat Sütçü , Muradiye Acar , Bülent Ediz

https://doi.org/10.26650/JARHS2024-1496234

Öz

Amaç: Bu çalışmanın amacı, hem gastrointestinal şikâyeti olan hastalarda bağırsak patojeni protozoonların prevalansını tespit ederek sürveyans verilerine katkı sağlamak hem de mikroskopi ve multipleks real-time PCR yöntemlerinin performanslarını karşılaştırmalı olarak değerlendirmektir.

Gereç ve Yöntem: Çalışmaya, en az bir gastrointestinal şikâyeti olan 40 yetişkin (18 yaş ve üzerinde) ve 40 çocuk (5-12 yaş arasında) gönüllü hasta dâhil edildi. Her hastadan günaşırı olarak 3 kez alınan gaita örnekleri; Nativ-Lugol, modifiye formol-eter çöktürme, Wheatley’in trikrom boyama ve modifiye Ziehl-Neelsen boyama teknikleri uygulandıktan sonra ışık mikroskobunda incelendi. Ayrıca, yalnızca ilk olarak alınan gaita örnekleri; DNA ekstraksiyonundan sonra, Allplex™ GI Parasite Assay paneli kullanılarak multipleks real-time PCR yöntemiyle test edildi. Yöntemlerin duyarlılıkları ve özgüllükleri arasındaki farklılıklar, Fisher’in kesin testi ile analiz edildi. p değerinin <0,05 olması, istatistiksel olarak anlamlı kabul edildi.

Bulgular: Bağırsak patojeni protozoonların prevalansının yetişkin grubunda %25, çocuk grubunda ise %35 olduğu tespit edildi. Yöntemlerin duyarlılıkları ve özgüllükleri arasındaki farklılıklar, istatistiksel olarak anlamlı bulunmadı (p=0,999).

Sonuç: Allplex™ GI Parasite Assay paneli kullanılarak gerçekleştirilen multipleks real-time PCR yönteminin yalnızca ilk örnek test edildiğinde bile etkili olması; bu yöntemin, bağırsak patojeni protozoonların rutin tanısında kullanılabileceğini düşündürmektedir.

Anahtar Kelimeler

Patojen, bağırsak, protozoon, prevalans, mikroskopi, PCR

INVESTIGATION OF THE PREVALENCE OF PATHOGENIC INTESTINAL PROTOZOANS IN PATIENTS WITH GASTROINTESTINAL COMPLAINTS VIA MICROSCOPY AND MULTIPLEX REAL-TIME PCR

Öz

Objective: This study aimed to contribute to surveillance data by determining the prevalence of pathogenic intestinal protozoans in patients with gastrointestinal complaints and to comparatively evaluate the performance of microscopy and multiplex real-time PCR methods.

Materials and Methods: Forty adults (18 years or older) and 40 children (aged 5 to 12 years) who volunteered patients with at least one gastrointestinal complaint were included in the study. Stool samples collected three times every other day from each patient were examined under a light microscope using Native-Lugol, modified formol-ether concentration, Wheatley’s modified trichrome staining, and Modified Ziehl-Neelsen staining techniques. In addition, the first stool samples were tested using multiplex real-time PCR with the Allplex™ GI Parasite Assay panel after DNA extraction. Differences between the sensitivities and specificities between the methods were analysed using Fisher’s exact test. A p-value of <0.05 was considered statistically significant.

Results: The prevalence of pathogenic intestinal protozoans was 25% in adults and 35% in children. Differences in the sensitivities and specificities of the methods were not found to be statistically significant (p=0.999).

Conclusion: Multiplex real-time PCR method performed using the Allplex™ GI Parasite Assay panel was effective even when only the first sample tested, suggesting that this method would be used for the routine diagnosis of pathogenic intestinal protozoans.

Anahtar Kelimeler

Pathogen, intestinal, protozoon, prevalence, microscopy, PCR

Kaynakça

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