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白斑综合症病毒(WSSV)的宿主调查 总被引：24，自引：3，他引：24
用地高辛 (DIG)标记的WSSVDNA探针斑点杂交与原位杂交技术 ,在中国对虾、斑节对虾、南美白对虾、刀额新对虾、脊尾白虾、天津厚蟹、日本大眼蟹体内检测到了WSSV ,它们是WSSV的天然宿主 ;在经人工感染的哈氏美人虾、短脊鼓虾、克氏原螯虾、肉球近方蟹、滕壶体内检测到了WSSV ;在球形侧腕水母、病虾池的桡足类等浮游生物、卤虫无节幼体以及人工浸泡感染卤虫成体体内没有检测到WSSV。经原位杂交检测 ,虾类的甲壳下上皮、胃上皮、附肢、造血组织、鳃等组织器官均可被WSSV侵染 ,其中甲壳下上皮和鳃对WSSV敏感 ;蟹类的甲壳下上皮和鳃对WSSV敏感 ;在中国对虾、南美白对虾、脊尾白虾、注射感染的克氏原螯虾的精巢中 ,精荚的结缔组织细胞和血细胞呈阳性 ,在中国对虾、脊尾白虾以及注射感染的短脊鼓虾的卵巢中 ,结缔组织细胞和滤泡细胞被WSSV感染。 相似文献
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PCR法制备地高辛标记探针斑点杂交
检测白斑综合症病毒(WSSV) 总被引：2，自引：0，他引：2
用PCR法成功制备了DIG标记探针，探针长度为547bp，探针的产量为21.6ng/μL。此探针与随机引物合成探针检测样品灵敏度相近。用此探针核酸斑点杂交法检测了54尾中国对虾。结果表明此探针在对白斑综合症病毒的检测、对虾暴发性流行病的诊断等方面具有很高的应用价值。 相似文献
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Edwardsiella tarda has become one of the most important emerging pathogens in aquaculture industry.Therefore,a rapid,reproducible,and sensitive method for detection and quantification of this pathogen is needed urgently.To achieve this purpose,we developed a TaqMan-based real-time PCR assay for detection and quantification of E.tarda.The assay targets the hemolysin activator HlyB domain protein of E.tarda.Our optimized TaqMan assay is capable of detecting as little as 40 fg of genomic DNA per reaction.A standard curve was generated from the threshold cycle values(y) against log10(E.tarda genomic DNA concentration) as x.The intra-and inter-assay coefficient of variation(CV) values were less than 2.06% and 1.05% respectively,indicating that the assay had good reproducibility.This method is highly specific to E.tarda strains,as it shows no cross-reactivity to Edwardsiella ictaluri,a member of the same genus,or to nine other fish-pathogenic bacteria species belonging to three other genera.This sensitive and specific real-time PCR assay provides a valuable tool for diagnostic quantitation of E.tarda in clinical samples. 相似文献
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Edwardsiella tarda is a major pathogen in aquatic environments that can cause heavy economic losses. An improved method for quick and accurate detection of E. tarda by loop-mediated isothermal amplification (LAMP) with two additional loop primers was developed by targeting the EsrB gene (EsrB-LAMP). In this method, the Mg 2+ concentration, reaction temperature, and reaction time were optimized to 8 mmol/L, 61°C, and 40 min, respectively. The detection limit with the EsrB gene was as low as 10 copies, which is 100 times more sensitive than that of conventional polymerase chain reaction (PCR). The EsrB-LAMP assay was shown more sensitive and rapid than previously reported LAMP assays targeting the hemolysin gene (hemolysin-LAMP) for detection of E. tarda. The EsrB-LAMP was also highly specific to E. tarda and had no cross-reaction with 13 other strains of bacteria. The assay can be carried out in a simple heating device and the EsrB-LAMP products can be visually detected by adding fluorescent dye to the reaction mixture. Taken together, the improved EsrB-LAMP diagnostic protocol has the potential for detection of E. tarda from indoor and outdoor samples. 相似文献
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