Efficiency of Tilapia Lake Virus Detection in Nile Tilapia by Nested RT-PCR and Semi- Nested RT-PCR
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Abstract
There are several methods for detecting Tilapia Lake virus (TiLV). The most popular methods for detecting RNA viruses are nested RT-PCR and semi-nested RT-PCR. The primers currently used are varied and are often designed to be specific for viruses found in each area. This study aimed to develop a detection method for TiLV in tilapia by designing two pairs of TiLVspecific primers and then detecting TiLV by nested RT-PCR and semi-nested RT-PCR, which examined suspected tilapia samples from fish cages in the Tapi River, Nakhon Si Thammarat Province. The results showed that nested RT-PCR (PCR product size of 307 bp) and semi-nested RT-PCR 2 (PCR product size of 496 bp) gave more accurate results than semi-nested RT-PCR 1 (PCR product size of 600 bp) while nested RT-PCR was more sensitive than semi-nested RT-PCR 2. The sensitivity and specificity of the nested RT-PCR method were each 100%.
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