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猪繁殖与呼吸综合症病毒四川株分离、鉴定及其病原特性研究 被引量:12
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作者 程安春 汪铭书 +3 位作者 希尼尼根 袁桂萍 周毅 陈孝跃 《黑龙江畜牧兽医》 CAS 北大核心 2004年第9期15-18,共4页
从四川地区发病仔猪分离到 3株病毒 (SC1、SC2和SC3) ,电镜下可见病毒呈球形 ,有囊膜 ,病毒能在Marc 14 5细胞上生长并致细胞出现CPE ,直径约 5 5nm ,核酸为RNA ,不凝集猪、黄牛、犬、兔、豚鼠、鸡、鸭、鹅的红细胞 ;pH值 2 ,3,4 ,8,9,1... 从四川地区发病仔猪分离到 3株病毒 (SC1、SC2和SC3) ,电镜下可见病毒呈球形 ,有囊膜 ,病毒能在Marc 14 5细胞上生长并致细胞出现CPE ,直径约 5 5nm ,核酸为RNA ,不凝集猪、黄牛、犬、兔、豚鼠、鸡、鸭、鹅的红细胞 ;pH值 2 ,3,4 ,8,9,10处理 30min和 5 6℃处理 6 0min病毒即失去对Marc 14 5细胞的感染能力。经病毒中和实验、特异性单克隆免疫荧光抗体和特异性RTPCR鉴定表明分离毒为美洲型猪繁殖与呼吸综合症病毒 (PRRSV) ,分离病毒素回归 30日龄仔猪可出现高热和呼吸道症状。研究首次从病原学角度证实美洲型PRRSV在四川的存在 ,利用分离毒制备的疫苗具有良好的免疫原性。 展开更多
关键词 繁殖呼吸综合症 病毒四川株 分离 鉴定 病原性
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禽网状内皮增生病毒囊膜基因gp90的克隆表达及ELISA方法的初步建立 被引量:1
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作者 余冲 黄勇 +3 位作者 田明星 石敏 李敏 赵芳芳 《浙江大学学报(农业与生命科学版)》 CAS CSCD 北大核心 2011年第4期363-370,共8页
根据禽网状内皮组织增生症病毒(reticuloendotheliosis virus,REV)SNV株前基因组cDNA序列,经生物信息学分析,设计并合成引物,以四川分离株REV-SC1为模板,扩增囊膜基因gp90抗原性、亲水性较高的121~1 023 bp基因片段;将目的基因克隆至p... 根据禽网状内皮组织增生症病毒(reticuloendotheliosis virus,REV)SNV株前基因组cDNA序列,经生物信息学分析,设计并合成引物,以四川分离株REV-SC1为模板,扩增囊膜基因gp90抗原性、亲水性较高的121~1 023 bp基因片段;将目的基因克隆至pMD-19T载体,进行测序和比对;将基因片段按正确的阅读框架定向克隆至pET-32a(+),转化大肠杆菌BL21(DE3),以IPTG诱导表达,包涵体洗涤纯化,探讨以重组gp90蛋白为包被抗原初步建立检测REV的间接酶联免疫吸附试验(ELISA)方法.结果表明:1)REV-SC1株gp90核苷酸序列与在GenBank上已发表序列的同源性最高为99%,于231位、870位发生了2个碱基突变;2)SDS-PAGE分析表明表达蛋白分子质量约为45 ku,与预期分子质量大小相符;免疫印迹(Western-blot)结果表明重组蛋白具有生物学活性;3)间接ELISA检测方法的最佳抗原包被质量浓度为1.5 mg·mL-1,一抗稀释度为1∶320,临界值为0.126;特异性、敏感性试验结果良好.综上,本试验成功表达了REVgp90蛋白,并对其在ELISA检测方面的应用作了初步探讨. 展开更多
关键词 网状内皮组织增生症病毒四川分离 GP90基因 克隆表达 间接酶联免疫吸附试验
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Prevalence and patterns of drug-resistance mutations among HIV-1 patients infected with CRF07_BC strains in Sichuan province,China 被引量:1
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作者 Ling Su Xia Zhou +4 位作者 Dan Yuan Hong Yang Dongbing Wei Guangmin Qin Shu Liang 《Virologica Sinica》 SCIE CAS CSCD 2014年第4期237-241,共5页
Little information is available on the prevalence of drug-resistance mutations in patients harboring the human immunodeficiency virus type 1(HIV-1) circulating recombinant form(CRF)07_BC variant in Sichuan, China. Thi... Little information is available on the prevalence of drug-resistance mutations in patients harboring the human immunodeficiency virus type 1(HIV-1) circulating recombinant form(CRF)07_BC variant in Sichuan, China. This study examined 375 plasma samples from patients with HIV-1 who were infected with the CRF07_BC strain, including 104 drug-naive participants and 271 in whom antiretroviral therapy(ART) had failed. Only one participant in the drug-naive group had a drug-resistance mutation(M46L), compared with 31.73% of those in whom ART had failed. Further analysis showed that 19.56% of strains contained mutations conferring resistance to non-nucleoside reverse transcriptase inhibitors(NNRTIs) alone, 0.74% were resistant to nucleoside reverse transcriptase inhibitors(NRTIs) alone, and 11.44% were dual-resistant to both NRTIs and NNRTIs. The most common mutation in the ART-failure group was M184V(35.88%), K103N(45.01%), Y181C(17.33%), and G190S/A(15.88%). The percentages of HIV-1 strains resistant to lamivudine, emtricitabine, efavirenz, etravirine, and nevirapine were 10.70%, 10.70%, 28.04%, 7.75%, and 26.20%, respectively. To explore site variants possibly related to drug resistance, variations in the ancestor/consensus CRF07_BC sequences from the therapy-naive and ART-failure groups were compared, and seven mutations at six positions were identified as being significantly differently distributed between the two groups(p<0.05). Detailed sequence data will provide information on CRF07_BC genetic characterizations, and improve our understanding of antiretroviral susceptibility and the evolution of drug-resistance mutations. This will be valuable in developing and implementing local public-health approaches for HIV drug-resistance prevention and treatment. 展开更多
关键词 HIV-1 CRF07_BC drug resistance SICHUAN China
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