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对虾传染性皮下及造血组织坏死病毒(IHHNV) LAMP检测方法的建立
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Abstract:
本研究建立了一种检测对虾传染性皮下及造血组织坏死病毒(Infectious hypodermal and hematopoietic necrosis virus, IHHNV)的环介导等温扩增法(Loop-mediated isothermal amplification, LAMP)。根据GenBank中注册的IHHNV-VN基因序列设计出6条特异性引物，成功建立了IHHNV-LAMP检测方法。该方法检出限为6 copies/μl，且与对虾其它常见疫病均无交叉反应。采用LAMP和常规PCR方法对40份具有典型IHHNV感染症状的对虾样品进行检测，结果显示：LAMP方法的阳性检出率为97.5%，PCR方法的阳性检出率为85%，表明建立的IHHNV-LAMP方法不仅具有高灵敏度、高特异性而且还有较高的准确性，能够为对虾传染性皮下及造血组织坏死病的早期快速诊断提供技术支撑。
This study established a loop-mediated isothermal amplification (LAMP) method for detecting Infectious hypodermal and hematopoietic necrosis virus (IHHNV) in shrimp. Six specific primers were designed based on the registered IHHNV-VN gene sequence in GenBank, and a successful IHHNV-LAMP detection method was established. The detection limit of this method is 6 copies/μl. And there is no cross reaction with other common diseases of shrimp. 40 shrimp samples with typical symptoms of IHHNV infection were detected using LAMP and conventional PCR methods. The results showed that the positive detection rate of LAMP method was 97.5%, and the positive detection rate of PCR method was 85%. This indicates that the established IHHNV-LAMP method not only has high sensitivity and specificity, but also high accuracy, which could provide technical support for the early and rapid diagnosis of Infectious hypodermal and hematopoietic necrosis virus in shrimp.
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