| 核酸斑点杂交分析法检测对虾皮下及造血组织坏死杆状病毒(HHNBV) |
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| 引用本文： | 史成银,宋晓玲,黄倢,杨丛海.核酸斑点杂交分析法检测对虾皮下及造血组织坏死杆状病毒(HHNBV)[J].海洋与湖沼,1999,30(5):486-490. |
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| 作者姓名： | 史成银 宋晓玲 黄倢 杨丛海 |
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| 作者单位： | 中国水产科学研究院黄海水产研究所农业部海水增养殖病害与生态重点开放实验室!青岛，266071，中国水产科学研究院黄海水产研究所农业部海水增养殖病害与生态重点开放实验室!青岛，266071，中国水产科学研究院黄海水产研究所农业部海水增养殖病害与生态重点开放实验室!青岛，266071 |
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| 基金项目： | 国家“八五”科技攻关项目!85-021-03-06 |
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| 摘 要： | 于１９９５年７月在青岛晨阳对虾养殖场采集患当性流行病的中国对虾样品，提纯ＨＨＮＢＶ并研制了一组地高辛标记的ＨＨＮＢＶ核酸探针。采用核酸斑点杂交方法了此组核酸探针的灵敏度和特异性。结果表明，此组探针检测ＨＨＮＢＶＤＮＡ的灵敏度为６２．５ｐｇ，对４６９ｎｇ病虾胃中的病毒可以检出；此组探针与３．７５μｇ健康对虾组织和２ｎｇ健康对虾ＤＮＡ均无交叉反应，１９９７年，应用此组特异性核酸探针检测了发病虾池对虾及
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| 关 键 词： | 中国对虾 核酸探针 斑点杂交 杆状病毒 造血组织 |
| 收稿时间： | 1/4/1998 12:00:00 AM |
| 修稿时间： | 1998/9/21 0:00:00 |
DETECTION OF HYPODERMAL AND HEMATOPOIETIC NECROSIS BACULOVIRUS (HHNBV) BY DIG-DNA PROBE DOT-BLOT HYBRIDIZATION |
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| SHl Cheng-yin,SONG Xiao-ling,HUANG Jie and YANG Cong-hai.DETECTION OF HYPODERMAL AND HEMATOPOIETIC NECROSIS BACULOVIRUS (HHNBV) BY DIG-DNA PROBE DOT-BLOT HYBRIDIZATION[J].Oceanologia Et Limnologia Sinica,1999,30(5):486-490. |
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| Authors: | SHl Cheng-yin SONG Xiao-ling HUANG Jie and YANG Cong-hai |
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| Abstract: | Diseased shrimps, Peneaus chinensis, were collected during an occurrence of explosiveepidemic disease at a shrimp farm of Chengyang, Qingdao in July 1995. The hypodermal andhematopoietic necrosis baculovirus (HHNBV) was isolated from these shrimps. Two differentdigoxigenjn (DlG) Iabeled DNA probes were constructed. The sensitivity and speciflcity of theseprobes were detected by DNA dot-blot hybridization assay. The results show that the sensitivitywas62.5pg to HHNBV DNA; these probes were able to hybridize with 469ng stomach hssues ofHHNBV-infected Penaeus chinensis, but not with HHNBV-uninfected P. chinenslj hssues and DNA.These probes were used to detect the sdrimps in diseased ponds and an urgent harvest pond in1997. The detecting results were intensely posihve. In contrast to PCR method, DIG DNA probedotblot hybridizahon had less procedures, which only needs simpIe reagents and apparatus, candetect hundreds specimen at onetime, and there is no need to extract DNA from specimen. So itwas easy to use. We have made a kind of dot-blot hybridizahon kit for HHNBV. As probes wereeasily synthesized the method of DNA probe can spread more easily than anh-serum method.Compared with PCR method, the DNA probe method has a lower sensihvity but higher specificity.The sequences of HHNBV DNA in clone C12 and C44 were known. We have designed severalpajrs of PCR primers according of these sequences. So we can amplify HHNBV DNA segments inspecimen with PCR, then hybridize those products with DNA probe. The sensihvity would highlyincrease and the specificity remains well. So these DIG labeled DNA probes have a high-appliedvalue in detechng HHNBV, diagnosing explosive epidemic disease and specific pathogen resistant(SPR) shrimp breeding. |
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| Keywords: | Penaeus chinensis DNA probe Dot-blot hybridization Detection Baculovirus |
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