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牛轮状病毒微滴式数字RT-PCR检测方法的建立及初步应用
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作者 谢世斌 郑辉 +6 位作者 沙洲 房琳琳 尼博 张慧 董雅琴 魏荣 崔进 《中国草食动物科学》 北大核心 2025年第1期44-49,共6页
本研究旨在建立更加灵敏的牛轮状病毒(Bovine rotavirus,BRV)检测方法,为BRV的病原学调查及畜牧养殖行业疫病防控提供更加准确和灵敏的技术支持。根据BRV VP6基因组序列,利用Oligo7.56软件设计并合成特异性引物与探针,通过优化反应条件... 本研究旨在建立更加灵敏的牛轮状病毒(Bovine rotavirus,BRV)检测方法,为BRV的病原学调查及畜牧养殖行业疫病防控提供更加准确和灵敏的技术支持。根据BRV VP6基因组序列,利用Oligo7.56软件设计并合成特异性引物与探针,通过优化反应条件,建立检测BRV的微滴式数字RT-PCR方法,同时开展特异性、敏感性和重复性试验,并将建立的方法应用于14份临床样品的检测。结果表明,本研究所建方法的最低检测限为1.83 copies/μL,具有良好的可重复性,与牛传染性鼻气管炎病毒、牛病毒性腹泻病毒、牛冠状病毒、牛呼吸道合胞体病毒和牛副流感病毒3型均无交叉反应。对14份临床样品进行检测,共检测出4份阳性样品,阳性率为28.57%,检测结果与实时荧光定量RT-PCR检测结果的符合率为100.00%。综上说明,本研究建立的BRV微滴式数字RT-PCR检测方法具有良好的特异性、敏感性和重复性,可应用于临床样本的检测,为BRV的诊断和流行病学调查提供了技术支持。 展开更多
关键词 牛轮状病毒 VP6基因 微滴式数字rt-pcr
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Activatable fluorescent probes for imaging and diagnosis of rheumatoid arthritis
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作者 Pan Luo Fu-Qiang Gao +5 位作者 Wei Sun Jun-You Li Cheng Wang Qing-Yu Zhang Zhi-Zhuo Li Peng Xu 《Military Medical Research》 SCIE CAS CSCD 2024年第2期287-307,共21页
Rheumatoid arthritis(RA)is a systemic autoimmune disease that is primarily manifested as synovitis and polyarticular opacity and typically leads to serious joint damage and irreversible disability,thus adversely affec... Rheumatoid arthritis(RA)is a systemic autoimmune disease that is primarily manifested as synovitis and polyarticular opacity and typically leads to serious joint damage and irreversible disability,thus adversely affecting locomotion ability and life quality.Consequently,good prognosis heavily relies on the early diagnosis and effective therapeutic monitoring of RA.Activatable fluorescent probes play vital roles in the detection and imaging of biomarkers for disease diagnosis and in vivo imaging.Herein,we review the fluorescent probes developed for the detection and imaging of RA biomarkers,namely reactive oxygen/nitrogen species(hypochlorous acid,peroxynitrite,hydroxyl radical,nitroxyl),pH,and cysteine,and address the related challenges and prospects to inspire the design of novel fluorescent probes and the improvement of their performance in RA studies. 展开更多
关键词 Rheumatoid arthritis fluorescent probe IMAGING DIAGNOSIS BIOMARKER
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Metal-organic cage as fluorescent probe for LiPF_(6)in lithium batteries
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作者 Xi Li Dehua Xu +3 位作者 Aoxuan Wang Chengxin Peng Xingjiang Liu Jiayan Luo 《Green Energy & Environment》 SCIE EI CAS CSCD 2024年第10期1592-1600,共9页
Lithium hexafluorophosphate(LiPF_(6)),the most commonly used lithium battery electrolyte salt,is vulnerable to heat and humidity.Quantitative and qualitative determination the variation of LiPF_(6)have always relied o... Lithium hexafluorophosphate(LiPF_(6)),the most commonly used lithium battery electrolyte salt,is vulnerable to heat and humidity.Quantitative and qualitative determination the variation of LiPF_(6)have always relied on advanced equipment.Herein,we develop a fast,convenient,high-selective fluorescence detection method based on metal-organic cages(MOC),whose emission is enhanced by nearly 20 times in the presence of LiPF_(6)with good stability and photobleaching resistance.The fluorescent probe can also detect moisture in battery electrolyte.We propose and verify that the luminescence enhancement is due to the presence of hydrogen bond-induced enhanced emission effect in cages.Fluorescent excitation-emission matrix spectra and variable-temperature nuclear magnetic resonance spectroscopy are employed to clarify the role of hydrogen bonds in guest-loaded cages.Density functional theory(DFT)calculation is applied to simulate the structure of host-guest complexes and estimate the adsorption energy involved in the system.The precisely matched lock-and-key model paves a new way for designing and fabricating novel host structures,enabling specific recognition of other target compounds. 展开更多
关键词 Metal-organic cage Selective fluorescence probe Moisture detection Hydrogen bond Host-guest complex
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TaqMan real-time fluorescent quantitative RT-PCR in detection of macrophage inflammatory protein-2γ mRNA in myocarditis murine
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作者 杨佳荟 沈茜 《Journal of Medical Colleges of PLA(China)》 CAS 2003年第5期301-304,共4页
Objective: To study the role of macrophage inflammatory protein (MIP)-2γ in myocarditis pathogenesis in BALB/c mice. Methods: The relationship between the progression of Coxsarckie virus B3(CVB3) viral myocarditis an... Objective: To study the role of macrophage inflammatory protein (MIP)-2γ in myocarditis pathogenesis in BALB/c mice. Methods: The relationship between the progression of Coxsarckie virus B3(CVB3) viral myocarditis and experimental autoimmune myocarditis and MIP-2γ mRNA expression in mouse was studied by TaqMan real-time fluorescent quantitative RT-PCR. Results: MIP-2γ mRNA expression rose on 3 to 5 d after CVB3 infection, reached peak on 7 d, and returned to normal level until 14 d, which corresponded well with the disease course. The MIP-2γ mRNA expression level rose significantly on the day 18 d after immunization with porcine cardiac myosin, which was consistent with pathological examination. Conclusion: MIP-2γ may be involved in the pathogenesis of myocarditis. 展开更多
关键词 TAQMAN real-time fluorescent quantitative rt-pcr MYOCARDITIS MIP-2γ MRNA
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A ratiometric fluorescent probe for hypoxanthine detection in aquatic products based on the enzyme mimics and fluorescence of cobalt-doped carbon nitride
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作者 Xin Wang Chengyi Hong +1 位作者 Zhengzhong Lin Zhiyong Huang 《Food Science and Human Wellness》 SCIE CSCD 2024年第2期879-884,共6页
A ratiometric fluorescent probe for hypoxanthine(Hx)detection was established based on the mimic enzyme and fluorescence characteristics of cobalt-doped graphite-phase carbon nitride(Co doped g-C_(3)N_(4)).In addition... A ratiometric fluorescent probe for hypoxanthine(Hx)detection was established based on the mimic enzyme and fluorescence characteristics of cobalt-doped graphite-phase carbon nitride(Co doped g-C_(3)N_(4)).In addition to emitting strong fluorescence,the peroxidase activity of Co doped g-C_(3)N_(4)can catalyze the reaction of O-phenylenediamine and H_(2)O_(2)to produce diallyl phthalate which can emit yellow fluorescence at 570 nm.Through the decomposition of Hx by xanthine oxidase,Hx can be indirectly detected by the generating hydrogen peroxide based on the measurement of fluorescent ratio I(F_(570)/F_(370)).The linear range was 1.7-272.2 mg/kg(R^(2)=0.997),and the detection limit was 1.52 mg/kg(3σ/K,n=9).The established method was applied to Hx detection in bass,grass carp,and shrimp,and the data were verified by HPLC.The result shows that the established probe is sensitive,accurate,and reliable,and can be used for Hx detection in aquatic products. 展开更多
关键词 Co doped g-C_(3)N_(4) Ratiometric fluorescent probe HYPOXANTHINE FRESHNESS Aquatic products
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Complexation and Fluorescence Enhancement of Bay-site Carboxyl Modified Perylene Dimide with Calcium Ion
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作者 GU Dandan ZHAO Haoru +4 位作者 JIANG Xiaoze QI Haohan LIAO Zuogui CHEN Jia SUN Bin 《发光学报》 北大核心 2025年第3期474-485,共12页
Bay-site carboxyl functionalized perylene diimide derivative 1,7-COOH-PDI-C_(12)(PDI-COOH)was synthesized and distinct enhanced fluorescence was observed through combining with calcium ion(Ca^(2+))in THF/H_(2)O soluti... Bay-site carboxyl functionalized perylene diimide derivative 1,7-COOH-PDI-C_(12)(PDI-COOH)was synthesized and distinct enhanced fluorescence was observed through combining with calcium ion(Ca^(2+))in THF/H_(2)O solution.The assembly and fluorescence behavior of PDI-COOH/Ca^(2+)were studied in detail by changing hydration state with different concentrations.Based on the differences in assembly morphology and stoichiometric ratios of PDICOOH/Ca^(2+),we proposed the fluorescence emission mechanism of PDI-COOH/Ca^(2+)in THF/H_(2)O and THF,respectively.This work reveals a novel strategy of aggregated state fluorescence enhancement and reminds us of the important role of water in molecular fluorescence emission and assembly. 展开更多
关键词 carboxyl functionalized fluorescence behavior perylene diimide photoinduced electron transfer effect calcium ion
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蓝舌病新疆分离株VP7蛋白编码基因序列分析及一步法RT-PCR方法的建立
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作者 马晓菁 谷文喜 +5 位作者 叶锋 刘帅 刘丽娅 谢彩云 钟旗 易新萍 《新疆农业科学》 CAS CSCD 北大核心 2024年第1期253-259,共7页
【目的】分析蓝舌病新疆分离株编码VP7蛋白S7基因保守序列,建立蓝舌病新疆分离株一步法RT-PCR方法,为新疆BTV分子流行病学调查及防控提供技术支持。【方法】采用二代测序技术获得新疆分离株S7基因序列,登陆GenBank对序列进行同源性Blas... 【目的】分析蓝舌病新疆分离株编码VP7蛋白S7基因保守序列,建立蓝舌病新疆分离株一步法RT-PCR方法,为新疆BTV分子流行病学调查及防控提供技术支持。【方法】采用二代测序技术获得新疆分离株S7基因序列,登陆GenBank对序列进行同源性Blast比对,用软件MEGA 5.0分析序列差异,根据蓝舌病新疆分离株编码VP7蛋白S7基因保守序列,运用软件Oligo6.0设计引物,对蓝舌病新疆分离株S7基因进行RT-PCR扩增,并验证该方法的特异性及敏感性。【结果】蓝舌病中国新疆分离株编码VP7蛋白S7基因序列与哈尔滨报道BTV分离株S7基因片段相似度较高为89.64%,与中国云南报道BTV-29型分离株、蒙古国BTV分离株S7基因片段相似度分别为87.49%和87.39%;与德国BTV分离株S7基因片段相似度为80.70%,其余均无同源性序列。中国新疆分离株S7基因片段序列与4条同源序列间存在26处核苷酸差异,9处氨基酸差异。【结论】建立的蓝舌病中国新疆分离株S7基因片段一步法RT-PCR方法具有良好的特异性,仅BTV中国新疆分离株扩增获得目的条带,该方法的敏感性为4.0×10^(3)copies/mL。 展开更多
关键词 蓝舌病 VP7蛋白 rt-pcr
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Photostable and Biocompatible Fluorescent Silicon Nanoparticles for Imaging-Guided Co-Delivery of siRNA and Doxorubicin to Drug-Resistant Cancer Cells 被引量:5
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作者 Daoxia Guo Xiaoyuan Ji +4 位作者 Fei Peng Yiling Zhong Binbin Chu Yuanyuan Su Yao He 《Nano-Micro Letters》 SCIE EI CAS CSCD 2019年第2期127-139,共13页
The development of effective and safe vehicles to deliver small interfering RNA(siRNA) and chemotherapeutics remains a major challenge in RNA interference-based combination therapy with chemotherapeutics,which has eme... The development of effective and safe vehicles to deliver small interfering RNA(siRNA) and chemotherapeutics remains a major challenge in RNA interference-based combination therapy with chemotherapeutics,which has emerged as a powerful platform to treat drug-resistant cancer cells.Herein,we describe the development of novel all-in-one fluorescent silicon nanoparticles(SiNPs)-based nanomedicine platform for imaging-guided co-delivery of siRNA and doxorubicin(DOX).This approach enhanced therapeutic efficacy in multidrug-resistant breast cancer cells(i.e.,MCF-7/ADR cells).Typically,the SiNP-based nanocarriers enhanced the stability of siRNA in a biological environment(i.e.,medium or RNase A) and imparted the responsive release behavior of siRNA,resulting in approximately 80% down-regulation of P-glycoprotein expression.Co-delivery of P-glycoprotein siRNA and DOX led to>35-fold decrease in the half maximal inhibitory concentration of DOX in comparison with free DOX,indicating the pronounced therapeutic efficiency of the resultant nanocomposites for drug-resistant breast cancer cells.The intracellular time-dependent release behaviors of siRNA and DOX were revealed through tracking the strong and stable fluorescence of SiNPs.These data provide valuable information for designing effective RNA interference-based co-delivery carriers. 展开更多
关键词 fluorescent silicon nanoparticles Drug resistance Gene therapy BIOIMAGING
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Super-strong and Intrinsically Fluorescent Silkworm Silk from Carbon Nanodots Feeding 被引量:5
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作者 Suna Fan Xiaoting Zheng +7 位作者 Qi Zhan Huihui Zhang Huili Shao Jiexin Wang Chengbo Cao Meifang Zhu Dan Wang Yaopeng Zhang 《Nano-Micro Letters》 SCIE EI CAS CSCD 2019年第4期326-336,共11页
Fluorescent silk is fundamentally important for the development of future tissue engineering scaffolds.Despite great progress in the preparation of a variety of colored silks,fluorescent silk with enhanced mechanical ... Fluorescent silk is fundamentally important for the development of future tissue engineering scaffolds.Despite great progress in the preparation of a variety of colored silks,fluorescent silk with enhanced mechanical properties has yet to be explored.In this study,we report on the fabrication of intrinsically super-strong fluorescent silk by feeding Bombyx mori silkworm carbon nanodots(CNDs).The CNDs were incorporated into silk fibroin,hindering the conformation transformation,confining crystallization,and inducing orientation of mesophase.The resultant silk exhibited super-strong mechanical properties with breaking strength of 521.9±82.7 MPa and breaking elongation of 19.2±4.3%,improvements of 55.1%and 53.6%,respectively,in comparison with regular silk.The CNDs-reinforced silk displayed intrinsic blue fluorescence when exposed to 405 nm laser and exhibited no cytotoxic effect on cells,suggesting that multi-functional silks would be potentially useful in bioimaging and other applications. 展开更多
关键词 SILKWORM SILK CARBON NANODOT Mechanical property fluorescent SILK FEEDING method
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In Situ Synthesis of Fluorescent Mesoporous Silica–Carbon Dot Nanohybrids Featuring Folate Receptor-Overexpressing Cancer Cell Targeting and Drug Delivery 被引量:5
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作者 Shuai Zhao Shan Sun +6 位作者 Kai Jiang Yuhui Wang Yu Liu Song Wu Zhongjun Li Qinghai Shu Hengwei Lin 《Nano-Micro Letters》 SCIE EI CAS CSCD 2019年第2期193-205,共13页
Multifunctional nanocarrier-based theranostics is supposed to overcome some key problems in cancer treatment.In this work,a novel method for the preparation of a fluorescent mesoporous silica–carbon dot nanohybrid wa... Multifunctional nanocarrier-based theranostics is supposed to overcome some key problems in cancer treatment.In this work,a novel method for the preparation of a fluorescent mesoporous silica–carbon dot nanohybrid was developed.Carbon dots(CDs),from folic acid as the raw material,were prepared in situ and anchored on the surface of amino-modified mesoporous silica nanoparticles(MSNs–NH2) via a microwave-assisted solvothermal reaction.The as-prepared nanohybrid(designated MSNs–CDs) not only exhibited strong and stable yellow emission but also preserved the unique features of MSNs(e.g.,mesoporous structure,large specific surface area,and good biocompatibility),demonstrating a potential capability for fluorescence imagingguided drug delivery.More interestingly,the MSNs–CDs nanohybrid was able to selectively target folate receptor-overexpressing cancer cells(e.g.,HeLa),indicating that folic acid still retained its function even after undergoing the solvothermal reaction.Benefited by these excellent properties,the fluorescent MSNs–CDs nanohybrid can be employed as a fluorescence-guided nanocarrier for the targeted deliveryof anticancer drugs(e.g.,doxorubicin),thereby enhancing chemotherapeutic efficacy and reducing side effects.Our studies may provide a facile strategy for the fabrication of multifunctional MSN-based theranostic platforms,which is beneficial in the diagnosis and therapy of cancers in future. 展开更多
关键词 MESOPOROUS silica nanoparticles CARBON DOTS fluorescence imaging Targeted drug delivery Chemotherapy
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Fluorescent Silicon Nanorods-Based Nanotheranostic Agents for Multimodal Imaging-Guided Photothermal Therapy 被引量:5
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作者 Mingyue Cui Sangmo Liu +5 位作者 Bin Song Daoxia Guo Jinhua Wang Guyue Hu Yuanyuan Su Yao He 《Nano-Micro Letters》 SCIE EI CAS CSCD 2019年第4期293-307,共15页
The utilization of diagnosis to guide/aid therapy procedures has shown great prospects in the era of personalized medicine along with the recognition of tumor heterogeneity and complexity.Herein,a kind of multifunctio... The utilization of diagnosis to guide/aid therapy procedures has shown great prospects in the era of personalized medicine along with the recognition of tumor heterogeneity and complexity.Herein,a kind of multifunctional silicon-based nanostructure,i.e.,gold nanoparticles-decorated fluorescent silicon nanorods(Au@SiNRs),is fabricated and exploited for tumor-targeted multimodal imaging-guided photothermal therapy.In particular,the prepared Au@SiNRs feature high photothermal conversion efficiency(~43.9%)and strong photothermal stability(photothermal performance stays constant after five-cycle NIR laser irradiation),making them high-performance agents for simultaneously photoacoustic and infrared thermal imaging.The Au@SiNRs are readily modified with targeting peptide ligands,enabling an enhanced tumor accumulation with a high value of^8.74%ID g?1.Taking advantages of these unique merits,the Au@SiNRs are superbly suitable for specifically ablating tumors in vivo without appreciable toxicity under the guidance of multimodal imaging.Typically,all the mice treated with the Au@SiNRs remain alive,and no distinct tumor recurrence is observed during 60-day investigation. 展开更多
关键词 Gold NANOPARTICLE fluorescent SILICON NANORODS Nanotheranostic MULTIMODAL imaging PHOTOTHERMAL therapy Tumor target
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帕利亚姆病毒实时荧光定量RT-PCR检测方法的建立与应用 被引量:1
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作者 杨恒 李占鸿 +5 位作者 宋子昂 高林 李卓然 廖德芳 肖雷 李华春 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第1期395-400,共6页
本研究拟建立帕利亚姆病毒(Palyam virus,PALV)血清型特异性实时荧光定量RT-PCR(qRT-PCR)方法用于临床样本或媒介中PALV血清型鉴定。根据我国流行PALV毒株的基因节段2序列,设计扩增引物和TaqMan探针,建立PALV血清型特异型qRT-PCR方法,... 本研究拟建立帕利亚姆病毒(Palyam virus,PALV)血清型特异性实时荧光定量RT-PCR(qRT-PCR)方法用于临床样本或媒介中PALV血清型鉴定。根据我国流行PALV毒株的基因节段2序列,设计扩增引物和TaqMan探针,建立PALV血清型特异型qRT-PCR方法,对方法的特异性、灵敏性与重复性进行评估;以我国分离的28株PALV和90份核酸阳性血液样本评估检测方法的可靠性;利用建立的方法对采集库蠓样本中携带的PALV进行血清型鉴定。结果显示,建立的PALV血清型qRT-PCR检测方法具有良好的特异性与灵敏性,可检出核酸拷贝数下限在22至28 copies·μL^(-1)。对28株PALV的qRT-PCR检测结果与病毒测序鉴定结果一致;对PALV不同感染阶段哨兵动物血液(90份)中的qRT-PCR鉴定结果与分离病毒的血清型鉴定结果一致;建立的方法可准确鉴定库蠓中携带PALV的血清型。本研究建立的PALV血清型qRT-PCR定型方法具有良好的特异强、敏感性与重复性,可用于PALV感染动物与媒介中PALV血清型的鉴定,具有良好的应用价值。 展开更多
关键词 帕利亚姆病毒 血清型鉴定 实时荧光定量rt-pcr 检测方法
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山羊地方性鼻内肿瘤病毒TaqMan荧光定量RT-PCR检测方法的建立及应用 被引量:2
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作者 李鹏飞 高桂琴 +6 位作者 周广青 吴锦艳 颜新敏 曹小安 何继军 袁莉刚 尚佑军 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第5期2259-2266,共8页
山羊地方性鼻内肿瘤(enzootic nasal tumor, ENT)是山羊的病毒性传染病,由山羊地方性鼻内肿瘤病毒(enzootic nasal tumor virus of goats, ENTV-2)所引起,感染后可以导致山羊的鼻道筛骨上皮细胞发生不可逆的癌变,已在世界范围内广泛存在... 山羊地方性鼻内肿瘤(enzootic nasal tumor, ENT)是山羊的病毒性传染病,由山羊地方性鼻内肿瘤病毒(enzootic nasal tumor virus of goats, ENTV-2)所引起,感染后可以导致山羊的鼻道筛骨上皮细胞发生不可逆的癌变,已在世界范围内广泛存在,对山羊养殖业造成严重的经济损失。为建立快速、准确且能定量分析ENTV-2的检测方法,本研究根据GenBank上发布的ENTV-2 env基因(MT254061.1)保守区域设计引物和TaqMan探针,建立ENTV-2 TaqMan探针荧光定量RT-PCR检测方法,并对其特异性、灵敏性、重复性与临床检测效果进行验证。结果显示,重组质粒标准品模板浓度为6.30×10^(2)~6.30×10^(7) copies·μL^(-1)呈现良好的线性关系,相关系数R^(2)为0.996 5,扩增效率为110%,线性方程的斜率为-2.953,最低检测限度为6.30×10^(1) copies·μL^(-1);组内变异系数和组间变异系数均小于2%,重复性好;与口蹄疫病毒((foot-and-mouth disease virus, FMDV)、小反刍兽疫病毒(peste des petits ruminants virus, PPRV)、蓝舌病毒(bluetongue virus)、羊内源性逆转录病毒(endogenous retroviruses)等均无交叉反应,表明该方法具有很好的特异性。利用本研究所建立的TaqMan探针实时荧光定量RT-PCR检测方法对29份临床样品进行检测,该方法较普通PCR方法的检出率更高。综上表明,本研究成功建立了ENTV-2 TaqMan探针实时荧光定量RT-PCR检测方法,为ENTV-2的快速诊断和流行病学调查提供技术手段。 展开更多
关键词 山羊地方性鼻内肿瘤病毒 羊内源性逆转录病毒 TAQMAN 荧光定量rt-pcr
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鸽微RNA病毒实时荧光定量RT-PCR检测方法的建立
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作者 张靖鹏 陈翠腾 +5 位作者 林琳 付环茹 李兆龙 江斌 黄瑜 万春和 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第2期860-866,共7页
旨在建立鸽微RNA病毒(pigeon megrivirus,PiMeV)实时荧光定量RT-PCR检测方法。本研究根据GenBank中PiMeVs序列特征设计特异性检测引物,从信鸽粪便中检测到PiMeV阳性(命名为PiMeV-CHN001株),并对其3 C基因进行核苷酸同源性比较和遗传进... 旨在建立鸽微RNA病毒(pigeon megrivirus,PiMeV)实时荧光定量RT-PCR检测方法。本研究根据GenBank中PiMeVs序列特征设计特异性检测引物,从信鸽粪便中检测到PiMeV阳性(命名为PiMeV-CHN001株),并对其3 C基因进行核苷酸同源性比较和遗传进化分析,明确其基因特征后,设计特异性实时荧光定量RT-PCR检测(RT-qPCR)引物组,建立检测PiMeV的RT-qPCR方法。结果显示:PiMeV-CHN001株3 C基因全长为591 bp,编码197个氨基酸,和其他2株野鸽源PiMeV(MeV-B1株和MeV-B2株)核苷酸相似性分别为89.5%和92.0%。建立的检测PiMeV的RT-qPCR方法的标准曲线Y轴截距为37.93,斜率为-3.335,相关系数为1.00,扩增效率为99.4%。特异性强,仅PiMeV出现特异性扩增信号和特异性峰值[Tm值为(81.69±0.22)℃],对鸽源禽流感病毒(avian influenza virus,AIV)、鸽源禽I型副黏病毒(pigeon paramyxovirus type I,PPMV-1)、鸽输血传播病毒(pigeon torque teno virus,PTTV)、鸽腺病毒(pigeon adenovirus,PiAd)及鸽圆环病毒(pigeon circovirus,PiCV)检测均未见特异性扩增信号;敏感性优,最低检测限为54.0拷贝·μL^(-1);重复性好,批内和批间变异系数均低于1.5%。用建立的检测方法对42份信鸽粪便样品进行检测,发现2份阳性样品(阳性率为4.76%)。本研究首次证实我国大陆地区信鸽中存在PiMeV,丰富了PiMeV宿主谱信息;建立的RT-qPCR方法为后续开展PiMeV流行病学研究提供支撑。 展开更多
关键词 信鸽 鸽微RNA病毒 3 C基因 序列分析 实时荧光定量rt-pcr方法
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Computational design of ratiometric two-photon fluorescent Zn2+probes based on quinoline and di-2-picolylamine moieties 被引量:1
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作者 Zhe Shao Wen-Ying Zhang Ke Zhao 《Chinese Physics B》 SCIE EI CAS CSCD 2022年第5期253-260,共8页
To improve two-photon absorption(TPA)response of a newly synthesized probe,a series of ratiometric two-photon fluorescent Zn^(2+) sensors based on quinoline and DPA moieties have been designed.The one-photon absorptio... To improve two-photon absorption(TPA)response of a newly synthesized probe,a series of ratiometric two-photon fluorescent Zn^(2+) sensors based on quinoline and DPA moieties have been designed.The one-photon absorption,TPA,and emission properties of the experimental and designed probes before and after coordination with Zn^(2+) are investigated employing the density functional theory in combination with response functions.The design consists of two levels.In the first level of design,five probes are constructed through using several electron acceptors or donors to increase accepting or donating ability of the fluorophores.It shows that all the designed probes have stronger TPA intensities at longer wavelengths with respect to the experimental probe because of the increased intra-molecular charge transfer.Moreover,it is found that the probe 4 built by adding an acyl unit has the largest TPA cross section among the designed structures due to the form of longer conjugated length and more linear backbone.One dimethylamino terminal attached along the skeleton can improve TPA intensity more efficiently than two side amino groups.Therefore,in the second level of design,a new probe 7 is formed by both an acyl unit and a dimethylamino terminal.It exhibits that the TPA cross sections of probe 7 and its zinc complex increase dramatically.Furthermore,the fluorescence quantum yields of the designed probes4 and 7 are calculated in a new way,which makes use of the relation between the computed difference of dipole moment and the measured fluorescence quantum yield.The result shows that our design also improves the fluorescence quantum yield considerably.All in all,the designed probes 4 and 7 not only possess enhanced TPA intensities but also have large differences of emission wavelength upon Zn^(2+) coordination and strong fluorescence intensity,which demonstrates that they are potential ratiometric two-photon fluorescent probes. 展开更多
关键词 two-photon absorption fluorescent probe zinc ion intra-molecular charge transfer
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Preparation and performance of fluorescent polyacrylamide microspheres as a profile control and tracer agent 被引量:1
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作者 Wan-Li Kang Lei-Lei Hu +3 位作者 Xiang-Feng Zhang Run-Mei Yang Hai-Ming Fan Jie Geng 《Petroleum Science》 SCIE CAS CSCD 2015年第3期483-491,共9页
Polyacrylamide microspheres have been suc- cessfully used to reduce water production in reservoirs, but it is impossible to distinguish polyacrylamide microspheres from polyacrylamide that is used to enhance oil recov... Polyacrylamide microspheres have been suc- cessfully used to reduce water production in reservoirs, but it is impossible to distinguish polyacrylamide microspheres from polyacrylamide that is used to enhance oil recovery and is already present in production fluids. In order to detect polyacrylamide microspheres in the reservoir pro- duced fluid, fluorescent polyacrylamide microspheres P(AM-BA-AMCO), which fluoresce under ultraviolet irradiation, were synthesized via an inverse suspension polymerization. In order to keep the particle size distribu- tion in a narrow range, the synthesis conditions of the polymerization were studied, including the stirring speed and the concentrations of initiator, NaaCO3, and dispersant. The bonding characteristics of microspheres were deter- mined by Fourier transform infrared spectroscopy. The surface morphology of these microspheres was observed under ultraviolet irradiation with an inverse fluorescence microscope. A laboratory evaluation test showed that the fluorescent polymer microspheres had good water swelling capability, thus they had the ability to plug and migrate in a sand pack. The plugging rate was 99.8 % and the residual resistance coefficient was 800 after microsphere treatment in the sand pack. Furthermore, the fluorescent microspheres and their fragments were accurately detected under ultra- violet irradiation in the produced fluid, even though theyhad experienced extrusion and deformation in the sand pack. 展开更多
关键词 Inverse suspension polymerization fluorescence Polyacrylamide microsphere Narrow sizedistribution - Profile control performance
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猪Linda病毒巢式RT-PCR检测方法的建立
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作者 于浩洋 王彩霞 +5 位作者 仇松寅 刘晓飞 景宏丽 吴绍强 冯春燕 林祥梅 《中国动物检疫》 CAS 2024年第1期95-101,共7页
猪Linda病毒能引起仔猪先天性震颤,严重危害养猪业的发展,目前在我国暂未发现感染病例。本研究根据猪Linda病毒Core-E~(ms)基因序列,设计并合成巢式RT-PCR引物,通过对退火温度、引物浓度等进行优化,建立了猪Linda病毒巢式RT-PCR检测方... 猪Linda病毒能引起仔猪先天性震颤,严重危害养猪业的发展,目前在我国暂未发现感染病例。本研究根据猪Linda病毒Core-E~(ms)基因序列,设计并合成巢式RT-PCR引物,通过对退火温度、引物浓度等进行优化,建立了猪Linda病毒巢式RT-PCR检测方法。结果显示,该方法具有良好的特异性,与非洲猪瘟病毒、猪瘟病毒、猪繁殖与呼吸综合征病毒、猪伪狂犬病病毒、猪圆环病毒2型均无交叉反应;敏感性好,对慢病毒阳性对照品的最低检出限为10~1 copies/μL,比普通RT-PCR灵敏10倍。使用该方法检测模拟病毒样品,发现最低检出限为10~1 TU/mL,与荧光定量RT-PCR方法一致。本研究首次建立了可检测猪Linda病毒的巢式RT-PCR方法,其特异性强、灵敏度高,为在口岸以及条件有限的场地对猪Linda病毒进行精准且快速的检测提供了有效技术手段,也为防范Linda病毒传入提供了有力技术支撑。 展开更多
关键词 猪Linda病毒 巢式rt-pcr 检测方法 非典型瘟病毒
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韦塞尔斯布朗病病毒RT-PCR快速检测方法的建立及应用
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作者 王奕婷 张风荣 +5 位作者 冯之航 郁蕾 薛俊欣 陈翔 王艳 赵光伟 《中国动物检疫》 CAS 2024年第10期96-101,共6页
为实现对韦塞尔斯布朗病病毒(Wesselsbron virus,WSLV)的快速检测,根据WSLV NS5基因保守序列,设计特异性扩增引物和探针,建立WSLV的TaqMan荧光定量RT-PCR和RT-PCR检测方法,并验证方法的敏感性、特异性及临床可行性。结果显示:所建立的... 为实现对韦塞尔斯布朗病病毒(Wesselsbron virus,WSLV)的快速检测,根据WSLV NS5基因保守序列,设计特异性扩增引物和探针,建立WSLV的TaqMan荧光定量RT-PCR和RT-PCR检测方法,并验证方法的敏感性、特异性及临床可行性。结果显示:所建立的两种方法的核酸检测下限分别为10、100 copies/μL,对日本脑炎病毒、西尼罗病毒和坦布苏病毒核酸均无交叉反应;分别利用建立的两种检测方法对100份入境马匹样品进行检测,未检出WSLV阳性,与南非参考实验室提供的双重荧光RT-PCR检测方法结果一致。结果说明,所建的两种方法敏感性高、特异性强,可用于临床样本检测,为韦塞尔斯布朗病的出入境检验检疫、流行病学调查及防控提供了技术支撑。 展开更多
关键词 韦塞尔斯布朗病病毒 荧光定量rt-pcr rt-pcr 快速检测
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非洲马瘟病毒和西尼罗病毒双重TaqMan荧光定量RT-PCR检测方法的建立及应用
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作者 钱佳豪 刘丹 +8 位作者 周师众 张博源 高建帅 蒋卉 范学政 张广智 丁家波 王春凤 沈青春 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第12期5873-5879,共7页
本研究旨在建立一种可同时检测非洲马瘟病毒(African horse sickness virus,AHSV)和西尼罗病毒(West Nile virus,WNV)的双重荧光定量RT-PCR检测方法。根据AHSV VP 7基因序列和WNV Poly Protein基因的高度保守区域设计特异性引物和探针,... 本研究旨在建立一种可同时检测非洲马瘟病毒(African horse sickness virus,AHSV)和西尼罗病毒(West Nile virus,WNV)的双重荧光定量RT-PCR检测方法。根据AHSV VP 7基因序列和WNV Poly Protein基因的高度保守区域设计特异性引物和探针,优化反应条件及体系,绘制标准曲线,对其特异性、敏感性和重复性进行测定,并采用该方法对临床样品进行检测验证。结果显示:该方法能够同时检测AHSV和WNV,且特异性良好,与马动脉炎病毒、马疱疹病毒1型、马传染性贫血病毒、马腺疫链球菌、马流感病毒等马病核酸均无交叉反应;敏感性高,AHSV和WNV最低检测限均为10 copies·μL^(-1);组内变异系数为0.04%~0.93%,组间变异系数为0.17%~4.83%,重复性良好;使用该方法对30份马全血样品进行检测,结果均为阴性。本试验建立的检测方法对马属动物检疫、非洲马瘟和西尼罗热的监测与防控具有重要意义。 展开更多
关键词 双重荧光定量rt-pcr 非洲马瘟病毒 西尼罗病毒
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Expression of the Capsid Precursor Protein gene of Foot-and-mouth Disease Virus and Green Fluorescent Protein Gene in BHK-21 Cells Mediated by Retroviral Vector
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作者 LI Jiong LIU Yan-hong +4 位作者 AN Fang-lan LIU Jun-lin LIU Xiang-tao SHANG You-jun YIN Hong 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第S1期70-75,共6页
We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constr... We have constructed a retroviral vector mediated mammalian cell expression system of the capsid precursor protein of foot-and-mouth disease virus(FMDV).The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed by sequentially inserting capsid precursor protein gene(P1) of FMDV and enhanced green fluorescent protein gene(EGFP) into pBABEpuro.The recombinant retroviral vector and the pVSV-G plasmid were co-transfected into packaging cells(GP2-293) by liposomemediated transduction to produce the pseudovirus.The pseudovirus was used to infect BHK-21 cells and resistant cells were screened with puromycin.Green fluorescent proteins were observed by fluorescence microscopy and expression of the capsid precursor protein gene of FMDV was detected by indirect immunofluorescence.The recombinant retroviral vector pBABEpuro-P1-2A-EGFP was constructed successfully.The capsid precursor protein of FMDV and green fluorescent protein were expressed in BHK-21 cells.The mammalian cell expression system for the capsid precursor protein of FMDV has been constructed successfully,which lays the foundation of development of a FMDV subunit vaccine. 展开更多
关键词 retroviral vector FMDV capsid precursor protein gene green fluorescent protein gene BHK-21 cell
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