Patojen Prototheca Türlerinin Tespitinde Konvansiyonel PCR Yönteminin Duyarlılığının Değerlendirilmesi

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

Kaynakça

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