An improved method for the detection of white spot syndrome virus (WSSV) by single-tube nested deoxyribonucleic acid amplification and a detection kit utilizing the method.

The method of detection employs a nucleic acid amplification process using PCR by first amplifying an outer WSSV genomic DNA fragment that may be present in extracted DNA of clinical specimen using an "external" set of two oligonucleotide primers. Sensitivity and specificity are then increased through a second round of nucleic acid amplification of an internal WSSV DNA setgment located within the amplified outer WSSV genomic DNA fragment using an "internal" set of two oligonucleotide primers. The amplification process with these nested primers also reduces background and non-specific amplification. A detection kit the above incorporating the above is disclosed.

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