DETEKSI Vibrio parahaemolyticus MENGGUNAKAN MARKA GEN PirA PADA UDANG VANAME (Litopenaeus vannamei) DENGAN REAL TIME PCR

Ronald Kriston Sauttua Nainggolan, Munti Yuhana, Sukenda Sukenda, Woro Nur Endang Sariati

Abstract


Pengujian validitas deteksi Vibrio parahaemolyticus strain AHPND pada udang vaname (Litopenaeus vannamei) dengan metode real time PCR dilakukan melalui tahapan pengujian yang mencakup spesifisitas, sensitivitas, repitabilitas, reproduktivitas, dan uji lapang pada tambak-tambak udang vaname di Indonesia. Uji performa analitik menunjukkan bahwa metode real time PCR dengan penciri gen VpPirA spesifik dalam mendeteksi V. parahaemolyticus strain AHPD dan memiliki sensitivitas hingga 10 kopi µL-1. Nilai cut off metode qPCR primer VpPirA adalah di Ct 39. Metode real time PCR dengan primer spesifik VpPirA juga memiliki nilai repitabilitas dan reproduktivitas yang baik dalam mendeteksi V. parahaemolyticus strain AHPND. Semua pengujian performa analitik metode qPCR telah memenuhi syarat keberterimaan dan dapat digunakan sebagai metode pengujian untuk mendeteksi V. parahaemolyticus strain AHPND. Hasil uji lapang untuk seluruh sampel yang berasal dari tambak udang vaname menunjukkan hasil negatif terinfeksi V. parahaemolyticus strain AHPND. Studi ini menyarankan bahwa metode real time PCR efektif dan valid dalam mendeteksi V. parahaemolyticus strain AHPND.

Testing the validity of Vibrio parahaemolyticus AHPND (Vp AHPND) strain detection in whiteleg shrimp (Litopenaeus vannamei) by real-time PCR method was carried out through several stages including specificity, sensitivity, repeatability, reproducibility, and sampling field tests in shrimp ponds in Indonesia. Analytical performance test showed that the use of real-time PCR method with VpPirA primer was effective in detecting Vp AHPND strain and had a sensitivity of up to 10 copies µL-1. The cut off value qPCR method of the primer VpPirA is at Ct 39. The real-time PCR method using VpPirA primer also has good repeatability and reproducibility values in detecting Vp AHPND strain. All analytical performance testings of the qPCR method meet the standard requirements to detect Vp AHPND strain. The result of field tests for all whiteleg shrimp samples from the shrimp ponds revealed negative infection of Vp AHPND. This study suggests that the real time PCR is effective and valid in detecting V. parahaemolyticus AHPND strain.


Keywords


Vibrio parahaemolyticus strain AHPND; Litopenaeus vannamei; real time PCR; deteksi; Vibrio parahaemolyticus AHPND strain; Litopenaeus vannamei; real-time PCR; detection

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DOI: http://dx.doi.org/10.15578/jra.15.2.2020.111-119


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