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对虾传染性皮下及造血器官坏死病毒(IHHNV)可感染世界各地养殖对虾,给对虾养殖业造成严重经济损失。本实验首次采用实时定量PCR法对广西地区的84份凡纳滨对虾样品进行检测,同时以常规PCR检测作对照。实时定量PCR检测阳性率为79·8%,常规PCR检测阳性率为40·5%,表明广西地区养殖的凡纳滨对虾IHHNV的感染率较高。将二者检测均呈阳性的30份样品扩增产物进行序列分析测序,测序结果通过DNA STAR软件包进行分析,并通过NCBI Blast与GenBank中的序列进行比对。结果证明,测定的是IHHNV序列。30份样品的IHHNV序列很保守,可以分为4种类型,仅有两个碱基的位置发生变异。实时定量PCR检测IHHNV,快速、灵敏、准确,特异性好,可以作为检测对虾感染病毒的有效方法。
Infectious shrimp subcutaneous and hematopoietic necrosis virus (IHHNV) can affect shrimp farming around the world, causing serious economic losses shrimp farming. In this experiment, 84 samples of Litopenaeus vannamei from Guangxi area were detected by real-time quantitative PCR for the first time, and compared with routine PCR detection. The positive rate of real-time PCR was 79.8%, and the positive rate of routine PCR was 40.5%, which indicated that the IHHNV infection rates of cultured Litopenaeus vannamei in Guangxi were higher. The amplified products of 30 samples which were positive by both were sequenced and sequenced. The sequencing results were analyzed by DNA STAR package and compared with the sequences in GenBank by NCBI Blast. The results demonstrate that IHHNV sequences were determined. The IHHNV sequences of 30 samples are very conservative and can be divided into 4 types, with only two base positions mutated. Real-time quantitative PCR detection of IHHNV, fast, sensitive, accurate and specific, can be used as an effective method to detect virus infection in shrimp.