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  fluorescent signals
Changes in fluorescent signals of DSM were found to depend on the EPO concentration and physiological status of thymus cells.
      
The four specific oligonucleotide probes including the matched and the mismatched by the fluorescent target sequence gave obviously different hybridization fluorescent signals.
      
The mixed probes of each sample were then hybridized with 4,096 cDNA arrays (4,000 unique human cDNA sequences), and the fluorescent signals were scanned by ScanArray 3000 scanner (General Scanning, Inc.).
      
Following an enrichment of mutant cells by use of 9-(1'-pyrene)nonanol/ultraviolet irradiation (P9OH/UV) method, five peroxisome-defective mutants were isolated by pursuing the fluorescent signals from GFP.
      
The slides were studied with a fluorescence microscope and the percentages of positive nuclei with aberrant fluorescent signals were counted.
      
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Objective To identify the IS6110-restriction fragment length polymorphism(RFLP )DNA fingerprinting patterns of some clinical isolates of Mycobacterium tuberculosis (MTB) isolated from Ningxia, Beijing and Shanghai in recent years, and to observe their epidemiological characteristics.Methods Chromosomal DNA of MTB was digested with endonuclease PvuII, then electrophoresed on agarose gel plate and transferred capillarilly to a nylon filter and hybridized with 245 bp fragment of IS6110 which labeled using...

Objective To identify the IS6110-restriction fragment length polymorphism(RFLP )DNA fingerprinting patterns of some clinical isolates of Mycobacterium tuberculosis (MTB) isolated from Ningxia, Beijing and Shanghai in recent years, and to observe their epidemiological characteristics.Methods Chromosomal DNA of MTB was digested with endonuclease PvuII, then electrophoresed on agarose gel plate and transferred capillarilly to a nylon filter and hybridized with 245 bp fragment of IS6110 which labeled using random primer fluorescein labeling kit.The RFLP patterns of Southern hybridization were inspected autofluorographically and the chromosomal DNAs of MTB were thereby typed.Results Most of 103 isolates of MTB shared 8~21 copies. Some of them were clustered. Strains isolated from Ningxia and Beijing had similar DNA fingerprinting patterns . One zero copy strain and one single copy strain were found among isolates from Shanghai.Conclusions IS6110-RFLP based typing is feasible for MTB molecular epidemiological study in China. Most of isolates of MTB show analyzable patterns. Isolates of MTB from Ningxia and Beijing have close genomic relation.

目的 建立宁夏、北京和上海等地结核分支杆菌临床分离株IS6 110 RFLPDNA指纹图谱 ,观察其流行病学特征。方法 提取结核分支杆菌基因组DNA ,经限制性内切酶PvuⅡ切割、琼脂糖凝胶电泳和Southern转印后 ,用荧光标记的IS6 110DNA序列中 2 45bp探针杂交 ,以核酸化学发光试剂盒探测荧光信号 ,比较各菌株指纹的IS6 110拷贝数和带型 ,分析不同地理区域流行菌株的特点。结果  10 3例结核患者的临床分离株 ,经 2 45bp探针杂交后的指纹图谱显示大部分菌株含 8~ 2 1个IS6 110拷贝。在宁夏和北京地区流行的结核分支杆菌带型具有共同特点 ,并有成簇分布的现象 ;在上海分离株中发现 1株零拷贝株和 1株单拷贝株。结论 IS6 110 RFLPDNA分型方法可用于我国流行的结核分支杆菌分子流行病学研究 ;宁夏分离株与北京分离株在基因上亲缘关系较相近

Objectives To study moleculer epidemiology of the strains of mycobacterium tuberculosis(MTB) isolated from Guyuan.Methods Chromosomal DNA of MTB was digested with endonuclease PvuII, then electrophresed on agarose gel plate and transferred capillarilly to a nylon filter and hybridized with 245bp fragment of IS6110 which labeled using random primer fluorescein labeling kit.The RFLP patterns of Southern hybridization were inspected autofluorographically and the chromosomal DNAs of MTB were thereby typed.Results...

Objectives To study moleculer epidemiology of the strains of mycobacterium tuberculosis(MTB) isolated from Guyuan.Methods Chromosomal DNA of MTB was digested with endonuclease PvuII, then electrophresed on agarose gel plate and transferred capillarilly to a nylon filter and hybridized with 245bp fragment of IS6110 which labeled using random primer fluorescein labeling kit.The RFLP patterns of Southern hybridization were inspected autofluorographically and the chromosomal DNAs of MTB were thereby typed.Results The strains of MTB isolated from Guyuan carry 10-22 copies,which are present in a less variety of chromosomal site.Conclusions There is low genotypic variability in Guyuan isolates.

目的 探讨宁夏固原地区结核分子流行病学的特点。方法 提取结核分枝杆菌基因组DNA ,经限制性内切酶PvuⅡ切割、琼脂糖凝胶电泳和Southern转印后 ,用荧光标记的IS6 110DNA序列中 2 45bp探针杂交 ,以核酸化学发光试剂盒探测荧光信号 ,比较各菌株指纹的IS6 110拷贝数和带型 ,分析流行菌株的特点。结果 固原地区流行结核分枝杆菌临床分离株DNA指纹图谱带型相似 ,IS6 110拷贝数较多。结论 固原地区流行的结核分枝杆菌多态性程度较低 ,在基因上具较近的亲缘关系。

Objective:To develop modified molecular beacon based fluorescence PCR assay for the detection of V. parahaemolyticus.The established method was appliedto the rapid diagnosis of V. parahaemolyticus food poisoning and sea food examination.Methods:Two sets of primers were selected from the core sequence of TDH gene publishedin GenBank,and the modified molecular beacon labeled at 5'end & 3'end with diffferent fluorophors was designed. The molecular beacon and primer set were tested against numerous strains from...

Objective:To develop modified molecular beacon based fluorescence PCR assay for the detection of V. parahaemolyticus.The established method was appliedto the rapid diagnosis of V. parahaemolyticus food poisoning and sea food examination.Methods:Two sets of primers were selected from the core sequence of TDH gene publishedin GenBank,and the modified molecular beacon labeled at 5'end & 3'end with diffferent fluorophors was designed. The molecular beacon and primer set were tested against numerous strains from 12 different bacterial species. Real-time PCR assay for V.parahaemolyticus was established. Then this assay was applied to the food poisoning diagnosis.Results:Only V. parahaemolyticus strains generated fluorescent signals, and no cross-reaction was observed with other 11 bacterias. The sensitivity achieved was 69cfu /ml or 6cfu/PCR reaction. The assay was used to detect 40 V. parahaemolyticus strains, no false signals were observed.Forty-eight food poisoning samples and 100 sea food samples were tested, nine samples were found V. parahaemolyticus positive by real time PCR. Among the tested positive samples, seven samples were detected positive by traditional culture method. The overall test could be finished within one day starting from sample preparation.Conclusion:The modified molecular beacon-based real-time PCR assay was rapid, sensitive, and specific.It required 2 hours for the detection. It could be applied to the rapid diagnosis of V. parahaemolyticu food poisoning & food microbiology examination.

目的 :建立改良分子信标 -实时 PCR检测副溶血弧菌的快速方法 ,应用于副溶血弧菌食物中毒的快速诊断和海产品检验。方法 :根据 Gen Bank公布副溶血弧菌的耐热直接溶血毒素基因 (TDH)的保守序列 ,设计一对引物和改良分子信标探针 ,用 FAM荧光剂标记探针的 5′,并进行特异性和灵敏度分析 ;同时以 11种细菌作对照 ,建立改良分子信标检测副溶血弧菌的实时 PCR反应体系 ,应用于副溶血弧菌食物中毒快速诊断和食品微生物检测。结果 :检测 12种细菌 ,只有副溶血弧菌有荧光信号 ,与其他细菌无交叉反应 ,DNA灵敏度为 16 6 .6 fg/ μl,菌液灵敏度为 6 9cfu/ ml或 6 cfu/ PCR反应体系。改良分子信标 -实时 PCR反应体系检测 4 0株副溶血弧菌均出现特异的荧光信号 ,无干扰。对 3起细菌性食物中毒共 4 8份样品和 10 0份海产品进行检测 ,9份副溶血弧菌实时 PCR阳性 ,其中 7份副溶血弧菌细菌培养阳性 ,其余样品都为阴性。检测时间仅需 2 h。结论 :改良分子信标 -实时 PCR检测体系快速、灵敏度高 ,特异性强 ,可用于副溶血弧菌食物中...

目的 :建立改良分子信标 -实时 PCR检测副溶血弧菌的快速方法 ,应用于副溶血弧菌食物中毒的快速诊断和海产品检验。方法 :根据 Gen Bank公布副溶血弧菌的耐热直接溶血毒素基因 (TDH)的保守序列 ,设计一对引物和改良分子信标探针 ,用 FAM荧光剂标记探针的 5′,并进行特异性和灵敏度分析 ;同时以 11种细菌作对照 ,建立改良分子信标检测副溶血弧菌的实时 PCR反应体系 ,应用于副溶血弧菌食物中毒快速诊断和食品微生物检测。结果 :检测 12种细菌 ,只有副溶血弧菌有荧光信号 ,与其他细菌无交叉反应 ,DNA灵敏度为 16 6 .6 fg/ μl,菌液灵敏度为 6 9cfu/ ml或 6 cfu/ PCR反应体系。改良分子信标 -实时 PCR反应体系检测 4 0株副溶血弧菌均出现特异的荧光信号 ,无干扰。对 3起细菌性食物中毒共 4 8份样品和 10 0份海产品进行检测 ,9份副溶血弧菌实时 PCR阳性 ,其中 7份副溶血弧菌细菌培养阳性 ,其余样品都为阴性。检测时间仅需 2 h。结论 :改良分子信标 -实时 PCR检测体系快速、灵敏度高 ,特异性强 ,可用于副溶血弧菌食物中毒的快速诊断 ,为食源性疾病的分子流行病学调查提供新的检测手段

 
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