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间接免疫荧光法检测栉孔扇贝急性病毒性坏死症病毒 被引量:3

Serological detection of acute viral necrobiotic virus (AVNV) in scallop Chlamys farreri and Argopecten irradians by indirect immunofluorescence(IIF)
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摘要 用纯化栉孔扇贝(Chlamysfarreri)急性病毒性坏死症病毒(AVNV)免疫兔子,以兔抗血清为一抗,荧光标记的羊抗兔抗体为二抗,采用冰冻切片技术,建立了栉孔扇贝AVNV的间接免疫荧光检测方法。用该方法分析栉孔扇贝和海湾扇贝(Argopectenirradians)体内AVNV感染强度,并对感染率进行统计。结果表明,栉孔扇贝的肾脏、肝胰腺中,AVNV阳性信号最强,呈现中度到重度感染,其AVNV感染率100%。鳃丝、性腺及闭壳肌中未检测到阳性信号。在海湾扇贝的肝胰腺及肾中AVNV的感染率也较高。提示AVNV感染扇贝的靶器官主要是肾脏、肝胰腺。对与栉孔扇贝同一海区养殖的海湾扇贝在栉孔扇贝发病期间存活率较高的原因进行了讨论。 A virus (acute viral necrosis virus,AVNV) was firstly discovered in the moribund Chlamys farreri sampled from the scallop farms along the coasts of Shandong Province during the period of high mortality,and the emergence of AVNV has been associated with mass death of C.farreri causing important economic losses.This severe scallop disease was temporarily named Acute Viral Necrobiotic Disease(AVND).Assays to detect AVNV in short time are necessary.In this study,a technique of detecting AVNV by indirect immunofluorescence(IIF) assay based on frozen section with polyclonal antibody is described.The samples including two- and three-year-old C.farreri,and bay scallop Argopecten irradians bred in the current year,cultured in Taipingjiao Gulf,Qingdao,Shandong Province,were collected after the burst of AVND in September 2003.Frozen section of main organ including gill,mantle,kidney,digestive gland,rear intestine,adductor muscle and gonad were prepared.The infection intensity of AVNV was detected by IIF assay,and infection rate was statistically analyzed.The results indicated that AVNV could infect gill,mantle,kidney,digestive gland and rear intestine of two-and three-year-old C.farreri,however,gonad and adductor muscle were negative,and the infection was the most serious in kidney and digestive gland with infection rate of 100%.The infection intensity was negative in mantle,and from negative to middle degree was found in gill and rear intestine in two-year-old C.farreri. The differences of infection intensity and infection rate between two- and three-year-old C.farreri were that three-year-old C.farreri had lower infection intensity and infection rate than that of two-year-old C.farreri,except the infection intensity in kidney of three-year-old C.farreri which was higher than that of two-year-old C.farreri (n=10).In Argopecten irradians,kidney,digestive gland,and mantle were also affected by AVNV at infection rate of 100%,80% and 10%,respectively (n=10).Furthermore,the infection intensity of each organ is quite different.The percentages of infection intensity of slight degree,middle degree and heavy degree in kidney were 30%,30%,and 40%,respectively (n=10),whereas only slight infection degree was found in mantle and digestive gland.Gill,rear intestine gonad and adductor muscle were negative to AVNV. Both C.farreri and A.irradians were the host of AVNV,and kidney and digestive gland of these two species were the main target tissues of AVNV.The reason why A.irradians kept healthy while massive death happened to C.farreri in summer was discussed.
出处 《中国水产科学》 CAS CSCD 北大核心 2005年第1期38-42,共5页 Journal of Fishery Sciences of China
基金 国家"973"课题项目资助(G1999012001) 山东省科技攻关项目(2004GG2205115).~~
关键词 栉孔扇贝 栉孔扇贝急性病毒性坏死症病毒 间接免疫荧光检测 Chlamys farreri Argopecten irradians acute viral necrobiotic virus(AVNV) indirect immunofluorescence(IIF)
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