Objective A RT-PCR method for detecting new goose astrovirus (NGAstV) was developed.
Method According to the ORF2 sequence, specific primers were designed according to Oligo 7, and the amplification conditions optimized for the new methodology.
Result The optimized reaction system applied 10 μL premix, 1 μL each of ORF2 upstream and downstream primer in the concentration of 10 μmol·L−1, 3 μL template, and sterile deionized water to make up 20 μL with the following testing steps: 94 ℃ for 2 min, 30 cycles of 94 ℃ for 30 s, 55 ℃ for 15 s, and 72 ℃ for 15 s, and kept at 72 ℃ for 10 min. The method amplified the specific fragment of NGAstV but no other common disease viruses of geese and delivered a detection sensitivity of 62 fg·μL−1 on the NGAstV nucleic acid. Out of 45 clinical samples, 15 were positively detected at a rate of 33.33% by the newly developed method.
Conclusion The new RT-PCR method exhibited the specificity and repeatability that could satisfactorily be used for detecting and/or epidemiological studies relating to NGAstV.
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