Patojen Prototheca Türlerinin Tespitinde Konvansiyonel PCR Yönteminin Duyarlılığının Değerlendirilmesi

Yıl 2025, Cilt: 29 Sayı: 3, 569 - 575, 25.12.2025

Enes Göksal , İnci Tüney

https://doi.org/10.19113/sdufenbed.1694623

Öz

Bu çalışmada, iki patojen Prototheca suşunun konvansiyonel PCR ve yeniden PCR (re-PCR) yöntemleriyle moleküler olarak tespit edilebilirlik sınırı (Limit of Detection, LOD) değerlendirilmiştir. P. ciferrii (SAG 2063) ve P. bovis (LZ-5) suşlarına ait DNA örnekleri, 1:2 oranında seri dilüsyonlara tabi tutulmuş ve PCR yöntemiyle değerlendirilmiştir. Konvansiyonel PCR ile P. ciferrii suşunda yaklaşık 9.1𝚡10-4 ng/µL, P. bovis suşunda ise yaklaşık 5.34𝚡10-6 ng/µL DNA konsantrasyonuna kadar amplifikasyon gözlenmiştir. Re-PCR uygulaması sayesinde her iki suşta da daha düşük DNA düzeylerinde bant elde edilmiştir. Bu veriler, Prototheca türlerinin çok düşük konsantrasyonlarda bile moleküler düzeyde saptanması için yeterli hassasiyete sahip olduğunu ve re-PCR uygulamasının tanısal duyarlılığı artırabileceğini göstermektedir.

Anahtar Kelimeler

Mikroalg , Tanısal eşik , Nadir patojenler , Çevresel tespit

Evaluating the Sensitivity of Conventional PCR for Detecting Pathogenic Prototheca Species

Öz

This study aimed to evaluate the molecular detection limit (Limit of Detection, LOD) of two pathogenic Prototheca strains using conventional PCR and re-amplification PCR (re-PCR) methods. Genomic DNA samples from Prototheca ciferrii (SAG 2063) and Prototheca bovis (LZ-5) were subjected to 1:2 serial dilutions and evaluated by PCR. In conventional PCR, amplification was observed up to approximately 9.1𝚡10-4 ng/µL for P. ciferrii and 5.34𝚡10-6 ng/µL for P. bovis. Re-PCR allowed the detection of amplification products at even lower DNA concentrations in both strains. These findings demonstrate that Prototheca species can be detected at very low DNA concentrations using molecular techniques and that re-PCR may enhance diagnostic sensitivity.

Anahtar Kelimeler

Microalgae , Diagnostic threshold , Rare pathogens , Environmental detection

Etik Beyan

In this study, we undertake that all the rules required to be followed within the scope of the "Higher Education Institutions Scientific Research and Publication Ethics Directive" are complied with, and that none of the actions stated under the heading "Actions Against Scientific Research and Publication Ethics" are not carried out.

Teşekkür

This article is extracted from Abdullah Enes GÖKSAL’s MSc. thesis entitled “Isolation and Characterization of Prototheca sp. from Environmental Samples”, 2025, İzmir, supervised by İnci TÜNEY.

Kaynakça

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