Comparison of RT-PCR and rRT-PCR Methods in Detection of Viral Hemorrhagic Septicemia Virus (VHSV) in Marine Ornamental Fish

¹ Department of Biology, Faculty of Science and Analytical Data, Institut Teknologi Sepuluh Nopember, 60111, Indonesia
² Indonesian Fish Quarantine Inspection Agency, Denpasar 80361, Bali, Indonesia

Abstract

Reverse Transcription PCR (RT-PCR) and Real Time Reverse Transcription PCR (rRT-PCR) are methods that can be used to detect VHSV (Viral Hemorrhagic Septicemia Virus) on freshwater and seawater fish. rRT-PCR is more widely used than the RT-PCR method because of several advantages such as requiring shorter time and fewer stages. Both methods have their own advantages and disadvantages, so it is necessary to compare the two methods in detecting the presence of VHSV in marine ornamental fish. The research was carried out with 3 steps: sensitivity test, specificity test, and sample test. The sensitivity test showed that the smallest dilution concentration that could be detected by both methods was the same at 10-3 with 100 copies in the 77bp band. Specificity tests on both methods showed that VHSV primers used are specific. Sample tests in both methods showed the same results in each of the samples tested. The results of sensitivity, specificity and sample tests conducted on RT-PCR and rRT-PCR methods showed the same results (positive) thus both methods can be used to detect the VHSV in marine ornamental fish.