共查询到18条相似文献，搜索用时 305 毫秒
1.
应用多重PCR检测鉴别对虾白斑综合征病毒和桃拉病毒 总被引：5，自引：0，他引：5
研究建立了一种可同时检测对虾白斑综合征病毒(WSSV)和桃拉病毒(TSV)的多重聚合酶链式反应(多重PCR)技术。根据WSSV和TSV基因序列，设计合成了2对分别与WSSV和TSV某段基因序列互补的引物，用这2对引物对同一样品中的WSSV DNA和TSV RNA模板进行多重PCR扩增。结果均同时得到了2条特异的与实验设计相符的306bp(WSSV)和231bp(TSV)多重PCR扩增带，而对其他对虾病病原的PCR扩增结果均为阴性；敏感性测定结果表明，该多重PCR技术能检出10pg的WSSV DNA和1pg的TSV RNA模板。 相似文献
2.
为了解广西沿海地区南美白对虾白斑综合症病毒(WSSV)的感染情况,本文采用野外采样和PCR检测方法调查钦州市、钦南区、港口区、防城区、东兴市、北海市、铁山港区和合浦县等8个监测区的南美白对虾WSSV感染情况.2013年共检测南美白对虾样本317份,WSSV总阳性率为19.6%.其中,虾苗和成虾的WSSV阳性率分别为1.5%和32.1%.在8个监测区中,北海市虾苗和成虾WSSV检出率均为最高,分别为5.6%和76.5%.总体上,2013年WSSV在广西沿海地区除铁山港外的南美白对虾养殖区均有不同程度的流行,且成虾平均阳性率高于虾苗. 相似文献
3.
据对虾白斑综合症病毒(WSSV)的基因保守序列,使用Primer Explorer V3软件设计了2条引物,利用PCR的扩增产物,结合变性高效液相色谱(DHPLC)技术,建立了一种对虾白斑综合症病毒的DHPLC快速检测方法。该检测方法特异性好,与传染性皮下和造血器官坏死病毒、斑节对虾杆状病毒、肝胰腺细小病毒、对虾杆状病毒以及对虾基因组DNA无交叉反应;检测灵敏度较高,约为10-5ng/μL,高于常规PCR及荧光定量PCR的灵敏度。用建立的DHPLC方法对100尾对虾样品进行了临床检测,结果与荧光定量PCR检测结果一致。WSSV的DHPLC检测方法,特异性强、灵敏度高,是对WSSV进行有效检测的一种新方法。 相似文献
4.
《中国预防兽医学报》2016,(4)
根据OIE《水生动物诊断手册》、农业部公告第683号和质检行业标准等规范性技术文件,为验证本实验室基于环介导恒温扩增技术(LAMP)研发的对虾白斑综合征病毒(WSSV)现场快速高灵敏度检测试剂盒的有效性,本研究对随机抽样的157套试剂盒的分析特异性(ASp)、分析灵敏度(ASe)、诊断特异性(DSp)、诊断灵敏度(DSe)、重复性和稳定性等进行分析和评价。ASp结果显示该试剂盒与黄头病毒、偷死野田村病毒、肝胰腺细小病毒、致急性肝胰腺坏死病副溶血弧菌、虾肝肠胞虫、传染性皮下及造血组织坏死病毒6种对虾病原及对虾组织核酸均无交叉反应;ASe为102拷贝/μL;与OIE标准中WSSV的套式PCR方法相比,该试剂盒测试374份临床样品的DSp为95.8%,DSe为94.6%;该试剂盒对阴性样品及强阳性样品的检测重复率为100%,弱阳性样品的检测重复率为88.9%;该试剂盒可以在-20℃保存6个月以上。所研制的WSSV现场快速高灵敏检测试剂盒与OIE标准套式PCR方法检测结果符合率高。本研究结果表明该试剂盒具有操作简便、快速、灵敏度高、特异性强、重复性好稳定性强等优点,能够满足实际应用中的WSSV高灵敏度检测需求。 相似文献
5.
为了解浙江省宁波市对虾主养区苗种虾肝肠胞虫(EHP)、虾血细胞虹彩病毒(SHIV)、对虾白斑综合征病毒(WSSV)以及导致急性肝胰腺坏死病的副溶血弧菌(VP)4种病原的携带情况及特点,2020年2—8月采用PCR检测方法,对象山、宁海、奉化、余姚、慈溪、鄞州等地虾苗进行4种病原检测。结果显示:共检测45个养殖场点的南美白对虾虾苗样品268批次,在41批次虾苗样品中检出至少1种病原,整体阳性率为15.30%;EHP阳性率最高(8.96%),其次是VP(2.98%)和SHIV(2.24%),WSSV最低(1.12%);产自浙江本省的虾苗阳性率最高(30.77%),产自广东(21.51%)、海南(11.54%)、福建(6.35%)的次之,产自山东的最低(0),经列联表分析方法计算χ2=13.757,表明虾苗病原携带率(阳性率)与苗种产地之间相关性极为显著;不同病原的携带率呈现不同的时间分布特征,但整体来看,6月病原阳性率相对较高,7—8月最低,均未检出。结果表明,宁波市南美白对虾虾苗存在一定程度的病原污染,其中EHP污染最为严重,产自浙江本省以及6月份的虾苗病原污染率最高,需要针对性加强防控。本研究为宁波市对虾养殖业的病害防控提供了参考。 相似文献
6.
《畜牧与兽医》2017,(7):79-82
根据鸡细小病毒(chicken parvovirus,ChPV)的保守基因NS1设计了1对特异性引物,建立并优化了能够快速检测ChPV的二温式PCR方法。结果表明:该二温式PCR只对ChPV敏感,扩增产物为302 bp的特异性条带,其最低能检测到38.6 fg的ChPV DNA;而对鸡新城疫病毒、H9亚型禽流感病毒、马立克氏病病毒、鸡传染性喉气管炎病毒、鸡传染性支气管炎病毒不敏感。对60份临床样品进行检测,检出率为20%(12/60)。说明所建立的ChPV二温式PCR方法是一种快速简便、特异性强、敏感性高的检测方法,适用于ChPV的临床检测。 相似文献
7.
为进一步研究对虾抗病毒免疫方法,使用实验室动物病毒感染死亡率分析,研究传染性皮下及造血器官坏死病毒(IHHNV)和白斑综合征病毒(WSSV)在感染南美白对虾后的相互作用机制。结果显示,预感染IHHNV后的对虾在进行WSSV攻毒后,对虾的存活率得到了提高。荧光PCR检测病毒量的结果显示,预感染IHHNV再用WSSV攻毒,存活虾体内的IHHNV含量比死亡虾高。研究结果初步证明,IHHNV和WSSV在对虾体内存在相互作用机制,且IHHNV具有抑制WSSV复制的作用。 相似文献
8.
《中国兽医杂志》2015,(10)
为建立快速、准确检测H3亚型禽流感病毒(AIV)的方法,根据GenBank中H3亚型AIV HA基因序列,设计出1对特异性引物,通过优化反应条件建立了H3亚型AIV二温式RT-PCR检测方法。对该法进行特异性、敏感性检测,并对256份临床样品进行检测。结果表明,该法只能扩增H3亚型AIV,对其他亚型AIV及常见禽病病原体不扩增;对H3亚型AIV检测下限为1×10~4拷贝/μL;256份临床样品检测结果与病毒分离鉴定结果一致。与普通RT-PCR相比该法节省了30min,表明所建立的H3亚型AIV二温式RT-PCR方法是一种快速、简便和特异的检测方法。 相似文献
9.
急性肝胰腺坏死病(AHPND)是一种近年新发现的南美白对虾疫病,其病原为一种特异的副溶血性弧菌(Vibrio parahemolyticus,VP)。本试验根据文献建立的PCR方法,对福建省9家规模化南美白对虾养殖场进行了AHPND检测,分析不同样本处理方式对检测结果的影响。从90份样本的27份中分离鉴定出81株VP,表明该地养殖场幼虾具有较高的VP感染风险;经PCR检测,在23份样本中,检出阳性菌株72株,其中16份样本的所有受检VP均为特异性菌株,6份样本的部分受检VP为特异性菌株,1份样本的所有受检VP均非特异性菌株,表明患病幼虾存在不同基因型VP同时感染情况;以肝胰腺DNA进行PCR检测,在90份样本中检出8份阳性,而以肝胰腺增菌液进行PCR检测,在90份样本中检出25份阳性,表明直接以肝胰腺DNA进行PCR检测的灵敏性较低。本研究为AHPND检测提供了技术支持。 相似文献
10.
广东省湛江地区南美白对虾桃拉综合征病原研究 总被引：1，自引：0，他引：1
桃拉综合征是由桃拉病毒（Taura syndromevirus，TSV）引起的，与白斑综合征病毒、皮下及造血组织坏死杆状病毒、黄头病毒一起被列为对虾养殖中危害最严重的四大病毒。本文对2005年广东省湛江地区部分养殖场的发病南美白对虾进行了组织病理学观察及病原的分子生物学检测，提供了桃拉病毒的显微结构图片，证实了本次广东省湛江地区部分养殖场南美白对虾急性传染病的主要病原之一是桃拉病毒，说明桃拉综合征在我国部分对虾养殖场仍存在着流行趋势。 相似文献
11.
Infectious hypodermal and hematopoietic necrosis virus is the causative agent of a shrimp disease which causes economic losses on a global scale. A pair of primers, I2814F/I3516R, was designed from the IHHNV genomic sequence (GenBank) that encodes for structural protein corresponding to nucleotides 2814-3516, which amplifies a 703 base pair (bp) region from the virus genome. PCR amplification with the primers generated a product of the expected size from the purified IHHNV DNA of Litopenaeus vannamei and IHHNV-infected penaeid populations but not from the IHHNV-free shrimp, white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV). The PCR amplicon described above was labeled with digoxigenin (DIG)-11-dUTP as a probe used for dot blot hybridization and in situ hybridization test. Under the optimized PCR conditions, the primers were detected by as little as 20 fg of purified IHHNV DNA, which contained only 8.83 x 10(3) copies of IHHNV, a 1000-fold greater than using dot blot hybridization. Sections of histopathology showed eosinophilic intranuclear inclusions (Cowdry type A inclusions or CAIs) in infected tissues while in situ hybridization, cells displayed an intense reaction with the DIG-labeled probe. PCR assay was developed to detect IHHNV in penaeid shrimp and other crustaceans from the rearing ponds of China (March 2001-June 2004). The positive rate was 51.5% (154 out of 299) and 8.3% (2 out of 24) for penaeid shrimp and crab samples, respectively. The survey demonstrated the presence of IHHNV in China. 相似文献
12.
Mauro A. Turriani M. Ioannoni A. Russo V. Martelli A. Di Giacinto O. Nardinocchi D. Berardinelli P. 《Veterinary research communications》2010,34(1):25-32
Challenge tests with Artemia four different development stages (nauplii, metanauplii, pseudoadults and adults) to white spot syndrome virus was carried
out by immersion challenge and virus-phytoplankton adhesion route in order to asses the possibility of Artemia acting as a vector of WSSV to penaeid shrimp Litopenaeus vannamei postlarvae. The WSSV succeeded in infecting four stages Artemia, and nested-PCR detection for WSSV revealed positive results to virus-phytoplankton adhesion route. No mass mortalities were
observed in penaeid shrimp postlarvae fed with WSSV-positive Artemia which exposed to WSSV by virus-phytoplankton adhesion route, whereas WSSV DNA detected in penaeid shrimp postlarvae by nested-PCR.
By contrary, no WSSV-positive was detected in any animal fed with WSSV-negative Artemia. These results indicated that Artemia could serve as a vector in WSSV transmission. 相似文献
13.
Christopher Marlowe A. Caipang Mary Paz N. Aguana 《Veterinary research communications》2010,34(7):597-605
Viral and bacterial pathogens have raised serious concerns in the sustainability of the shrimp culture industry in the Philippines.
Heavy mortality associated with luminous vibriosis and white spot syndrome virus (WSSV) infection has been the major problem
besetting the industry. Using published PCR protocols for the diagnosis of vibriosis and white spot syndrome virus (WSSV)
disease in shrimp, we optimized these assays that could be suited to the shrimp aquaculture setting in the Philippines. Genomic
DNAs of Vibrio spp. that exhibited luminescence as well as those that grew on thiosulfate citrate bile salt sucrose agar (TCBS) were used
for the PCR amplification of the ribonuclease P (RNase P) gene. There was differential amplification of the RNase P gene based
on the phenotypic characters of the Vibrio spp. Similar results were also obtained using direct colony PCR of the bacterial colonies. White spot syndrome virus was
also detected in the infected shrimp and there were differences in the detection frequency in relation to the tissues used
for PCR amplification. Duplex PCR was also optimized that could be used for simultaneous detection of these pathogens in shrimp. 相似文献
14.
15.
White spot syndrome virus (WSSV), a rod-shaped double-stranded DNA virus, is an infectious agent causing fatal disease in shrimp farming around the globe. Within shrimp populations WSSV is transmitted very fast, however, the modes and dynamics of transmission of this virus are not well understood. In the current study the dynamics of disease transmission of WSSV were investigated in small, closed populations of Penaeus monodon and Penaeus vannamei. Pair cohabitation experiments using PCR as a readout for virus infection were used to estimate transmission parameters for WSSV in these two species. The mortality rate of contact-infected shrimp in P. monodon was higher than the rate in P. vannamei. The transmission rate parameters for WSSV were not different between the two species. The relative contribution of direct and indirect transmission rates of WSSV differed between the two species. For P. vannamei the direct contact transmission rate of WSSV was significantly lower than the indirect environmental transmission rate, but for P. monodon, the opposite was found. The reproduction ratio R0 for WSSV for these two species of shrimp was estimated to be above one: 2.07 (95%CI 1.53, 2.79) for P. monodon and 1.51 (95%CI 1.12, 2.03) for P. vannamei. The difference in R0 between the two species is due to a lower host mortality and hence a longer infectious period of WSSV in P. monodon. 相似文献
16.
17.
对虾白斑综合征病毒亚单位疫苗研究进展 总被引：2，自引：1，他引：1
对虾白斑综合征病毒（White spot syndromevirus,WSSV）是一种可以引起养殖对虾暴发性死亡的传染性病原,由于其强烈的传染性和极高的致死率,使得人们在应对它时,必须侧重于早期的防控。近年的研究表明,对虾存在类免疫（quasi—immune）机制．而WSSV的重组蛋白可以诱导对虾产生抗病保护效应。目前有关对虾白斑综合征病毒亚单位疫苗的研究大多围绕诱导产生高效免疫应答能力的囊膜蛋白进行免疫接种方式来展开。文章对该领域研究成果做一综述,旨在为今后WSSV的防控提供参考。 相似文献
18.
克氏原螯虾口服疫苗免疫对白斑综合征病毒人工感染的保护作用 总被引：2，自引：0，他引：2
利用家蚕一杆状病毒表达系统在蚕蛹体内表达的重组病毒囊膜蛋白rVp28制成的疫苗，口服免疫克氏原螯虾，观察了其对白斑综合征病毒人工感染的保护作用。对试验虾分别进行了2％rVp28、2％普通蚕蛹液（阳性对照）和普通饲料（阴性对照）等3个处理，持续口服免疫75d。免疫35d后口服攻毒，20d内rVp28组的累积死亡率为36．67％，与阴性对照相比差异显著（P〈0．05），危疫保护率（PRP）达59．26％；注射攻毒后20d内．rVp28组的PRP为49．99％。免疫后55d对存活虾再口服攻毒，20d内rVp28组的累积死亡率为36．84％，与阴性对照相比差异显著（P〈0．05）．PRP为63．16％；2次注射攻毒后．rVp28组的PRP为31．25％。结果表明，重组囊膜蛋白rVp28免疫后．对口服感染具有显著的保护作用，对注射感染有一定的保护作用． 相似文献