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| 1 | 白斑综合症病毒实时荧光LAMP检测方法的建立及应用显示文摘研究利用ESE-Quant tube scanner检测平台,建立了一套基于环介导等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP)的实时荧光检测方法,用于白斑综合征病毒(White Spot Syndrome Virus,WSSV)的检测;并在此基础上,与巢式PCR、Real-time PCR和其他已发表的4种LAMP方法在检测灵敏度、实际应用方面进行比较。结果显示,研究建立的实时荧光LAMP检测方法在63℃恒温反应30min可检测到最低为105倍稀释的基因组DNA模板,与Real-time PCR检测方法的灵敏度相当,高于巢式PCR和其他已发表的4种LAMP方法的检测灵敏度;而且特异性较好,与传染性皮下及造血组织坏死病毒等5种常见对虾病原DNA均无交叉反应。通过构建质粒进一步进行灵敏度测试显示,本研究建立的实时荧光LAMP检测方法最低检测限度为24个拷贝质粒DNA,检出时间亦为30min。通过对66份待检样品的检测结果显示,实时荧光LAMP检测方法的检出阳性率为7.57%,准确率为100%,高于其他WSSV的检测方法。因此,研究建立的WSSV实时荧光LAMP检测方法,操作简单,反应速度快,特异性好,灵敏度高,成本低廉,可以直观、实时地观察反应的进行情况,适合对虾养殖现场及诊断实验室的WSSV快速检测。 | 李红梅 江晓 刘助红 赵哲 任春华 胡超群 石磊 王广军 | 2015 | 水生生物学报2015,39,1: | 15 |
| 2 | Effect of Different Diet Combinations on WSSV Load and Survival Rate of Fenneropenaeus chinensis Larvae显示文摘Using microalgae( SA),artificial feed( AF),Brachionus plicatilis( BP) and brine shrimp larvae( BS),four diets including SA + AF,SA + AF + BP,SA + AF + BS and SA + AF + BP + BS were formulated and used to rear Fenneropenaeus chinensis larvae. The fertilized eggs fed with SA + AF were divided into disinfection group with iodophor and untreated group,and the fertilized eggs fed with other diets were all disinfected with iodophor. The survival rates,body weight and WSSV load of post larvae at each stage were compared. The results showed that there was no significant difference in the survival rate of prawns at the N-Z stage by feeding with different diets( P > 0. 05),but the survival rate of prawns fed with BP was higher than others( P < 0. 01) at the Z-P stage. The body weight of post larvae( P10) fed with SA + AF + BP was less than that fed with SA + AF + BP + BS( P <0. 01). The WSSV load of prawns( 10. 52 ±3. 3 copies/ng DNA) fed with SA + AF were significantly lower than that of other treatments( P < 0. 05) at the P10 stage. During the P11- P60 stage,four kinds of diets,including Ruditapes philippinarum foot muscle( CF) + artificial feed( AF),R. philippinarum foot muscle radiated by60 Co γ-ray( RCF) + artificial feed( AF),potassium permanganate-disinfected R. philippinarum foot muscle( DCF) + artificial feed( AF) and artificial feed( AF),were used to feed prawns. The prawns fed with CF + AF gained the greatest increase of body weight and body length,but had no significant difference with that fed with AF( P > 0. 05); prawns fed with CF + AF and AF had significant difference with the other two groups( P < 0. 05). The survival rates of prawns fed with four different diets had no significant difference with each other( P > 0. 05). WSSV artificial infection test showed that the accumulated mortality of prawns fed with four diets were higher than 90%,and the difference was not significant( P > 0. 05). | Xiao Guangxia Meng Xianhong Kong Jie Cao Baoxiang Liu Ning Zhang Qingwen Luo Kun Li Zhanjun | 2015 | Animal Husbandry and Feed Science2015,7,6: | 0 |
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