Diagnostic potential of digital polymerase chain reaction for quantification of the protozoan parasite Perkinsus olseni in the Manila clam Ruditapes philippinarum
Seung-Hyeon Kim, Biyun Zhao and Kyung-Il Park
Department of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University
The purpose of this study was to test the potential of digital PCR (dPCR) for diagnosing Perkinsus olseni. dPCR is currently one of the most effective molecular diagnosis techniques, as it is capable of quantifying the target sequence without a standard. Here, a primer pair and a probe were developed in the non-transcribed spacer region for diagnosis of P. olseni. In the sensitivity and specificity test, dPCR was able to detect as little as one P. olseni trophozoite, and could discriminate P. olseni from P. marinus. The number of P. olseni cells measured by dPCR was very similar to that determined by conventional cell counting. In conclusion, this study confirms that P. olseni can be quantified using dPCR. Additional validation is needed to access the efficiency of the technique in host tissues or environmental samples.
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