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分别针对桃拉综合征病毒(TSV)基因、传染性皮下和造血器官坏死病毒(IHHNV)基因,设计了2对特异性引物,通过对多重RT-PCR扩增条件的优化,建立了快速检测TSV和IHHNV的二温式多重RT-PCR技术。特异性和敏感性试验结果表明,该技术只对TSV RNA和IHHNV DNA进行扩增,分别得到1条231bp的TSV特异性cDNA扩增片段和1条356bp的IHHNV特异性DNA扩增片段,对其它对虾病原核酸的扩增结果为阴性,该技术最低能同时检测到10pg的TSV RNA和IHHNV DNA,具有高度的特异性和敏感性。临床应用试验证明,该技术可以用于对虾养殖业中TSV和IHHNV的快速检测和鉴别诊断。
Two pairs of specific primers were designed for the gene of Taura syndrome virus (TSV), infectious subcutaneous and hematopoietic necrosis virus (IHHNV), respectively. Through the optimization of multiple RT-PCR amplification conditions, the rapid detection of TSV And IHHNV two temperature multiple RT-PCR technology. The results of specificity and sensitivity test showed that only TSV RNA and IHHNV DNA were amplified by this technique. One 231bp TSV-specific cDNA fragment and one 356bp IHHNV-specific DNA fragment were obtained, The result of amplification of shrimp pathogenic nucleic acid is negative, the technology can detect 10pg of TSV RNA and IHHNV DNA at the same time with low specificity, and has high specificity and sensitivity. The clinical application test proves that the technology can be used for rapid detection and differential diagnosis of TSV and IHHNV in shrimp farming.