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Fluorescent Detection of Succinylcholine via an Amide Naphthotube-Based Indicator Displacement Assay
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作者 Yin Ye Wang Hui +4 位作者 Wu Jianfang Wang Lili Yang Liupan Zhao Chengda Yao Huan 《有机化学》 北大核心 2025年第8期2953-2959,共7页
Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities.... Succinylcholine(SC)is a widely used depolarizing muscle relaxant,but improper use can lead to arrhythmias and,in severe cases,pose a life-threatening risk.Additionally,some criminals exploit SC for illicit activities.Therefore,rapid SC detection is paramount for clinical practice and public safety.Currently,however,limited methods are available for the rapid detection of SC.A fluorescent indicator displacement assay sensor based on molecular recognition of an amide naphthotube was developed.This sensor enabled the rapid fluorescent detection of SC through competitive binding between SC and methylene blue with the amide naphthotube.The sensor exhibited exceptional sensitivity with a detection limit as low as 1.1μmol/L and a detection range of 1.1~60μmol/L,coupled with outstanding selectivity and robust stability.Furthermore,this sensor accurately determined SC levels in biological samples such as serum.In summary,this research provides a new solution for the rapid and accurate sensing of SC in complex matrices and offers new insights for the swift identification and detection of toxins. 展开更多
关键词 SUCCINYLCHOLINE molecular recognition indicator displacement assay fluorescent sensor
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改良型PCR-RFLP法快速检测降糖药物基因多态性组合
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作者 翟振谦 饶梓雯 +3 位作者 孙良平 李力 龙玉舟 欧阳东成 《实验室检测》 2025年第22期13-15,共3页
目的建立一种聚合酶链式反应-限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)的改良方法,实现与降糖药物疗效及不良反应相关的基因多态性快速检测。方法以2型糖尿病(type 2 diabe... 目的建立一种聚合酶链式反应-限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)的改良方法,实现与降糖药物疗效及不良反应相关的基因多态性快速检测。方法以2型糖尿病(type 2 diabetes mellitus,T2DM)患者及健康对照的外周血DNA为研究对象,设计覆盖SLC22A1(rs628031、rs12208357)、SLC47A1(rs2289669)、KCNJ11[rs5219(E23K)]、ABCC8、CYP2C9(^(*)2、^(*)3)和TCF7L2(rs7903146)等位点的多重引物体系,引入错配引物并结合顺序/并联酶切,优化扩增与条带判读。结果建立的体系最低DNA投料量5~10 ng,特异性>98%,批内/批间RSD<5%,与Sanger测序一致率达99%以上。单个96孔板可完成约80例检测,单样本成本低于测序的20%。初步临床分析提示部分基因型与二甲双胍或磺脲类疗效存在差异。结论改良型多重PCR-RFLP体系可在单管或分池流程下实现多位点联合判型,方法学性能稳定、成本可控;可作为测序技术的筛查性补充工具,用于降糖药物相关基因多态性检测。 展开更多
关键词 改良型pcr-rflp 降糖药物 基因多态性 个体化用药 2型糖尿病
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An improved reporter gene assay for evaluating the biological activity of recombinant human growth hormone
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作者 Xiaoming Zhang Heyang Li +8 位作者 Ying Huang Ping Lv Lvyin Wang Kezheng Xu Yi Li Xinyue Hu Yue Sun Cheng-gang Liang Jing Li 《Journal of Pharmaceutical Analysis》 2025年第5期1160-1162,共3页
Recombinant human growth hormone(rhGH)has been widely used for the treatment of disorders associated with GH deficiency and multiple clinical indications[1].Accurate determination of biological activity is essential i... Recombinant human growth hormone(rhGH)has been widely used for the treatment of disorders associated with GH deficiency and multiple clinical indications[1].Accurate determination of biological activity is essential in the development,registration,and quality control of rhGH pharmaceutical products[2].However,the existing in vivo bioassay procedure based on somatropin-induced weight gain in rats is complicated,and the use of a rat cell line-based approach(Nb2-11 bioassay),which measures the production of adenosine triphosphate(ATP)as a direct indicator of cell growth,has a low mechanism of action(MOA)relevance.Therefore,novel rhGH bioassays are still needed.To this end,we developed a reporter gene assay(RGA)based on the GH/insulin-like growth factor-1(IGF-1)axis. 展开更多
关键词 determination biological activity nb bioassay vivo bioassay treatment disorders associated BIOassay recombinant human growth hormone rhgh reporter gene assay somatropin
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A one-pot Cas12i3/Cas13d-based assay for rapid portable identification of genotypes I and II African swine fever viruses
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作者 Zhe Wang Haili Wang +7 位作者 Xinghui Yan Jin Li Yu Wang Guosong Qin Wenbo Sun Yanfang Wang Gaiping Zhang Jianguo Zhao 《Journal of Integrative Agriculture》 2025年第11期4456-4459,共4页
African swine fever(ASF),caused by the African swine fever virus(ASFV),has brought enormous economic loss and represents a major threat to the global pig industry(Vergne et al.2017).ASFVs are divided into 24 genotypes... African swine fever(ASF),caused by the African swine fever virus(ASFV),has brought enormous economic loss and represents a major threat to the global pig industry(Vergne et al.2017).ASFVs are divided into 24 genotypes based on their B646L gene,with only genotypes Ⅰ and Ⅱ circulating globally(Dixon et al.2019). 展开更多
关键词 one pot african swine fever asf caused african swine fever virus asfv b l genewith assay cas d RAPID cas i
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结肠小袋纤毛虫PCR-RFLP分型方法的建立 被引量:1
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作者 徐啊慧 冯彩彩 +6 位作者 丰山旺 赵立卓 齐闻新 张雯 胡苏辉 王天奇 闫文朝 《中国人兽共患病学报》 CAS CSCD 北大核心 2024年第9期829-833,840,共6页
目的建立高效、特异的结肠小袋纤毛虫遗传亚型分析方法。方法选择限制性内切酶ApoI和PflMI对结肠小袋纤毛虫ITS1-5.8S rDNA-ITS2的PCR扩增产物进行酶切分析,建立PCR-RFLP分型方法,利用建立的PCR-RFLP方法对猪源、羊源和豚鼠源临床粪便... 目的建立高效、特异的结肠小袋纤毛虫遗传亚型分析方法。方法选择限制性内切酶ApoI和PflMI对结肠小袋纤毛虫ITS1-5.8S rDNA-ITS2的PCR扩增产物进行酶切分析,建立PCR-RFLP分型方法,利用建立的PCR-RFLP方法对猪源、羊源和豚鼠源临床粪便样品进行遗传亚型分析。结果基于ApoI和PflMI的PCR-RFLP方法可以准确区分结肠小袋纤毛虫遗传变异型A和B,用PflMI可以进一步将遗传变异型B细分为B-c和B-t两个亚型。与镜检和测序结果比较,建立的PCR-RFLP方法具有良好的特异性和更高的灵敏性,不仅可以鉴定临床样品中结肠小袋纤毛虫单个亚型,而且可以鉴别单个样品中结肠小袋纤毛虫多个亚型的混合感染。结论本研究成功建立了结肠小袋纤毛虫PCR-RFLP方法,可用于结肠小袋纤毛虫遗传多态性鉴定和分子流行病学研究。 展开更多
关键词 结肠小袋纤毛虫 pcr-rflp分析 豚鼠
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利用PCR-RFLP方法快速鉴别中国牛蒡属药用植物
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作者 黄彦昌 宋跃岳 +6 位作者 许亮 郑汉 董玉玮 张娜 胡传银 窦德强 康廷国 《中华中医药学刊》 CAS 北大核心 2024年第12期80-83,I0021,共5页
目的随着近十几年来国内学者对牛蒡属药用植物研究的深入,中国牛蒡属植物鉴定需要一种快速、准确的分子鉴别手段。因此研究拟通过利用聚合酶链式反应-限制性酶切长度多态性(polymerase chain reaction restriction frag-ment length pol... 目的随着近十几年来国内学者对牛蒡属药用植物研究的深入,中国牛蒡属植物鉴定需要一种快速、准确的分子鉴别手段。因此研究拟通过利用聚合酶链式反应-限制性酶切长度多态性(polymerase chain reaction restriction frag-ment length polymorphism,PCR-RFLP)建立一种快速鉴定中国牛蒡属药用植物的方法。方法通过对两种中国牛蒡属药用植物牛蒡与毛头牛蒡常用鉴定DNA条形码进行限制性核酸内切酶图谱分析,找到牛蒡在内源转录间隔区1(internal transcribed spacer-1,ITS1)片段中有一个单核苷酸多态性(Single-nucleotide polymorphism,SNP)位点,正好为BsaAⅠ酶(YAC/GTR)的酶切位点,根据该酶切位点,设计引物对目标片段进行扩增。另外建立PCR体系:95℃预变性5min,循环反应40次(90℃20s,60℃20s,72℃20s),72℃延伸5min,4℃保温。建立限制性内切酶BsaAⅠ酶切体系,制作琼脂糖凝胶电泳观察结果。结果电泳结果表明在经过BsaAⅠ酶切后牛蒡将会产生长度分别为101bp与125bp的短条带,而毛头牛蒡未被切开仍为226bp的长条带,成功将两种药用植物区分开。该方法简单、快速、准确,满足日常对中国牛蒡属植物的鉴定。结论该实验通过从牛蒡与毛头牛蒡的ITS1序列入手,利用PCR-RFLP技术首次完成了对两种植物的鉴别研究,建立了中国牛蒡属植物的PCR-RFLP鉴别方法。 展开更多
关键词 牛蒡 毛头牛蒡 鉴别 pcr-rflp
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千里香杂合度PCR-RFLP快速评估方法的建立
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作者 王博铖 陈梓媛 +3 位作者 华中一 田慧 谢文波 袁媛 《中国实验方剂学杂志》 CAS CSCD 北大核心 2024年第4期29-34,共6页
目的:建立一种快速评估千里香种质材料杂合度的方法,为制定千里香的优异种质繁育策略和促进种质创新提供依据。方法:以65株千里香重测序数据为基础,检测并筛选单核苷酸多态性(SNPs),利用其中20个SNP位点,通过自编脚本将其转化为限制性... 目的:建立一种快速评估千里香种质材料杂合度的方法,为制定千里香的优异种质繁育策略和促进种质创新提供依据。方法:以65株千里香重测序数据为基础,检测并筛选单核苷酸多态性(SNPs),利用其中20个SNP位点,通过自编脚本将其转化为限制性内切酶片段长度多态性(RFLP)标记;采用聚合酶链式反应-限制性内切酶长度多态性(PCR-RFLP)法对12份千里香种质的20个RFLP标记进行检测,根据RFLP标记位点酶切片段数目计算千里香种质的杂合度;采用plink软件计算12份千里香种质的全基因组杂合度,并比较不同种质杂合度评估方法得到的结果。结果:PCR-RFLP法和基因组重测序法计算的杂合度间差异无统计学意义。利用PCR-RFLP法筛选获得了8份杂合度<30%的种质材料,利用基因组重测序法筛选获得9份杂合度<30%的种质材料,2种方法计算杂合度均<30%的种质材料共7份。结论:该文建立了千里香种质杂合度评估的PCR-RFLP法,其精确度为87.5%,准确率为77.8%,该法可为其他药用植物种质杂合度评估方法研究提供借鉴。 展开更多
关键词 千里香 聚合酶链式反应-限制性内切酶长度多态性(pcr-rflp) 单核苷酸多态性(SNPs) 杂合度 全基因组重测序
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Ultra-highly selective recognition of nucleosides over nucleotides by rational modification of tetralactam macrocycle and its application in enzyme assay
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作者 Huan Yao Jian Qin +6 位作者 Yan-Fang Wang Song-Meng Wang Liu-Huan Yi Shi-Yao Li Fangfang Du Liu-Pan Yang Li-Li Wang 《Chinese Chemical Letters》 SCIE CAS CSCD 2024年第6期274-278,共5页
Artificial macrocycle with high binding selectivity in water is often challenging but urgently needed in various research and application areas.Herein,we report a new water-soluble biomimetic tetralactam macrocycle an... Artificial macrocycle with high binding selectivity in water is often challenging but urgently needed in various research and application areas.Herein,we report a new water-soluble biomimetic tetralactam macrocycle and realize the ultra-high selectivity to nucleosides over corresponding monophosphate nucleotides by rational modification.The introduction of charged groups at the periphery of endofunctionalized cavity makes the selectivity(guanosine to guanosine 5-monophosphate)increase remarkably from 100 to 1119.Based on the ultra-high selectivity of biomimetic tetralactam macrocycle,the sensitive CD73 enzyme activity assay was then achieved through product-selective fluorescence indicator displacement assay.Furthermore,the capability of the proposed method for inhibitor screening was successfully displayed. 展开更多
关键词 Rational modification Biomimetic macrocycle Ultra-high selectivity Enzyme assay
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一种基于线粒体COI PCR-RFLP单酶切快速鉴定4种巨蛎属牡蛎的方法
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作者 崔中望 于诗奇 +1 位作者 缪雄平 阙华勇 《海洋科学》 CAS CSCD 北大核心 2024年第8期68-73,共6页
本文报道了一种基于线粒体细胞色素氧化酶亚基I(mt COI)基因序列PCR-RFLP单酶切的牡蛎物种鉴定方法。本方法可快速鉴定福建牡蛎(Crassostrea angulata)、熊本牡蛎(Crassostrea sikamea)、香港牡蛎(Crassostrea hongkongensis)和近江牡蛎... 本文报道了一种基于线粒体细胞色素氧化酶亚基I(mt COI)基因序列PCR-RFLP单酶切的牡蛎物种鉴定方法。本方法可快速鉴定福建牡蛎(Crassostrea angulata)、熊本牡蛎(Crassostrea sikamea)、香港牡蛎(Crassostrea hongkongensis)和近江牡蛎(Crassostrea ariakensis)等中国沿海常见的4种巨蛎属(Crassostrea)牡蛎。该方法以甲基转移酶(Msp I)作为限制性内切酶,对4种巨蛎属牡蛎的线粒体DNA COI扩增序列进行酶切,以得到的特异性条带为依据进行物种鉴定。本方法的鉴定结果与COI测序方法的鉴定结果一致,并且筛选出的单一的限制性内切酶Msp I在4种牡蛎的COI序列中不存在酶切位点的突变,准确率达到100%,能够为巨蛎属牡蛎的物种鉴别提供简便可靠的技术支撑。 展开更多
关键词 巨蛎属(Crassostrea)牡蛎 线粒体COI pcr-rflp
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Reversed-Phase-HPLC Assay Method for Simultaneous Estimation of Sorbitol, Sodium Lactate, and Sodium Chlorides in Pharmaceutical Formulations and Drug Solution for Infusion
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作者 Sreenivas Pippalla Venugopal Komreddy +2 位作者 Srinivasulu Kasa Vaishnavi Chintala Poluri Venkata Reddy 《American Journal of Analytical Chemistry》 CAS 2024年第2期57-71,共15页
A rapid, straightforward, sensitive, efficient, and cost-effective reverse-phase high-performance liquid chromatographic method was employed for the simultaneous determination of Sorbitol, Sodium Lactate, and Chloride... A rapid, straightforward, sensitive, efficient, and cost-effective reverse-phase high-performance liquid chromatographic method was employed for the simultaneous determination of Sorbitol, Sodium Lactate, and Chlorides in a drug solution for infusion. Sorbitol, Sodium lactate, and Chloride are all officially recognized in the USP monograph. Assay methods are provided through various techniques, with titrations being ineffective for trace-level quantification. Alternatively, IC, AAS, and ICP-MS, though highly accurate, are costly and often unavailable to most testing facilities. When considering methods, it’s important to prioritize both quality control requirements and user-friendly techniques. A simple HPLC simultaneous method was developed for the quantification of Chlorides, Sorbitol, and Sodium Lactate with a shorter run time. The separation utilized a Shimpack SCR-102(H) ion exclusion analytical column (7.9 mm × 300 mm, 7 μm), with a flow rate of 0.6 mL per min. The column compartment temperature was maintained at 40°C, and the injection volume was set at 10 μL, with detection at 200 nm. All measurements were conducted in a 0.1% solution of phosphoric acid. The analytical curves demonstrated linearity (r > 0.9999) in the concentration range of 0.79 to 3.8 mg per mL for Sodium Lactate (SL), 0.16 to 0.79 mg per mL for Sodium Chloride (SC), and 1.5 to 7.2 mg per mL for Sorbitol. Validation of the developed method followed the guidelines of the International Conference on Harmonization (ICH Q2B) and USP. The method exhibited precision, robustness, accuracy, and selectivity. In accelerated stability testing over 6 months, no significant variations were observed in organoleptic analysis and pH. Consequently, the developed method is deemed suitable for routine quality control analyses, enabling the simultaneous determination of Sodium Lactate, Sodium Chloride, and Sorbitol in pharmaceutical formulations and infusions. 展开更多
关键词 SORBITOL Sodium Lactate and Chloride assay Analytical Validation HPLC
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Interferon-gamma release assays as a tool for differential diagnosis of gastrointestinal tuberculosis
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作者 Tsvetelina Velikova Anita Aleksandrova 《World Journal of Clinical Cases》 SCIE 2024年第27期6015-6019,共5页
In this editorial,we comment on an article published in a recent issue of the World Journal of Clinical Cases.There is a pressing need for reliable tools for diagnosing tuberculosis(TB)of the gastrointestinal tract.De... In this editorial,we comment on an article published in a recent issue of the World Journal of Clinical Cases.There is a pressing need for reliable tools for diagnosing tuberculosis(TB)of the gastrointestinal tract.Despite advancements in the diagnosis and treatment,TB remains a global health challenge.Ali et al demon-strated that TB may mimic gastrointestinal conditions,such as gastric outlet obstruction,causing a delay in the diagnosis.Furthermore,the latter complication is frequently observed during infections,including Helicobacter pylori,and rarely is related to TB,as in the presented case.In line with this,we think that laboratory tests based on interferon-gamma release assays can be a helpful tool for diagnosing latent TB paced in the gastrointestinal tract.Innovative strategies and approaches for diagnosing latent/active extra pulmonary TB are crucial for establishing the diagnosis early and enhancing treatment strategies to mitigate the global burden of TB. 展开更多
关键词 TUBERCULOSIS Gastrointestinal tuberculosis Interferon-gamma release assay IGRA Primary gastroduodenal tuberculosis Gastric outlet obstruction Case report
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Evaluation of Interferon-Gamma Release Assay Testing and Tuberculin Skin Test for Early Diagnosis of Tuberculosis in Children and Adolescents
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作者 Yelda Sorguç Miray Çelebi Yılmaz +4 位作者 Yüce Ayhan Yakup Yaman Şener Tulumoğlu Aybüke Akaslan Kara İlker Devrim 《Open Journal of Pediatrics》 2024年第3期558-567,共10页
Background: This study aimed to evaluate the diagnostic value of interferon-γ release assay (IGRA), a sensitive microbiological diagnostic method, in children and adolescents with suspected tuberculosis in a country ... Background: This study aimed to evaluate the diagnostic value of interferon-γ release assay (IGRA), a sensitive microbiological diagnostic method, in children and adolescents with suspected tuberculosis in a country with a high burden of tuberculosis. Method: This study included 581 children and adolescents aged 4 - 19 years who were suspected of having tuberculosis, were latently infected with Mycobacterium tuberculosis, and had received at least one dose of BCG vaccine between April 17, 2019, and February 24, 2021. The study evaluated the TST results of 106 patients who had a positive Quantiferon test and were suspected of having tuberculosis. Results: The study included 581 patients aged between 4 and 19 years. Of these, 106 patients tested positive for the Quantiferon test, while 19 were indeterminate and 456 were negative. The Quantiferon test positivity rate was 18.24%. Among the 106 QFT-Plus-positive cases, 23 patients also tested positive for TST. The difference in distribution was found to be statistically significant. Conclusion: The QFT-Plus test is considered an alternative to TST and other microbiological diagnostic methods for early tuberculosis diagnosis, particularly in children and adolescents. 展开更多
关键词 Interferon Gamma Release assay CHILDREN Tuberculin Test CHILDREN Latent Tuberculosis
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Characterization of Bulinus Snails in Sô-Ava and Azowlissè, Two Localities in Southern Benin, Using PCR-RFLP
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作者 Elokou Alabi Codjo Gaston Ouikoun +4 位作者 Halfane Lehmane Haziz Sina Michele Sezonlin Adolphe Adjanohoun Lamine Baba-Moussa 《Open Journal of Ecology》 2024年第12期938-949,共12页
Schistosomiasis is a public health concern in Benin. Freshwater snails of the genus Bulinus serve as intermediate hosts for schistosomes, trematode parasites responsible for bilharzia. The urinary form, caused by Schi... Schistosomiasis is a public health concern in Benin. Freshwater snails of the genus Bulinus serve as intermediate hosts for schistosomes, trematode parasites responsible for bilharzia. The urinary form, caused by Schistosoma haematobium, is the most widespread and is transmitted to humans by these mollusks, with Bulinus truncatus and Bulinus globosus being the most important species. Effective strategies to combat the transmission of these parasites require a prior understanding of the molecular characterization of Bulinus snails. For this purpose, 293 Bulinus snails were collected and morphologically identified from two localities in southern Benin, Sô-Ava and Azowlissè. The snails were preserved in absolute alcohol. To achieve the set objectives, DNA was extracted from the collected biological material, and SSU gene fragments were amplified. Using PCR-RFLP, the amplified fragments were digested with the restriction endonucleases HaeIII, HinfI, and DdeI to perform molecular characterization. In this study, 80 individuals of B. globosus and 10 of B. truncatus were subjected to molecular analysis. The PCR-RFLP profiles showed bands of different sizes for the Bulinus species when analyzed with the three endonucleases using the SSU molecular marker. PCR-RFLP analysis revealed that the snails belonged to the freshwater genus Bulinus, including Bulinus globosus and B. truncatus, based on reference profiles from studies conducted in Nigeria, which enabled precise identification of these gastropods. This study provided initial insights, although still incomplete, into the molecular diversity of these species. 展开更多
关键词 Bulinus globosus Truncatus Molecular Diversity pcr-rflp BENIN
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Interferon Gamma Release Assay for Latent Tuberculosis Screening in High TB-Endemic Region: A Retrospective Study
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作者 Shaheda Anwar Ahmed Abu Saleh +5 位作者 SM Ali Ahmed Saika Farook Farhan Muhib Udoy Maisha Islam Tanvir Ahmed Siddiqui Md Sultan Ahmed 《Journal of Tuberculosis Research》 2024年第4期183-192,共10页
Introduction: Tuberculosis (TB) remains a significant public health challenge, particularly in high-endemicity settings where latent TB infections (LTBI) contribute to ongoing transmission. Early identification and ma... Introduction: Tuberculosis (TB) remains a significant public health challenge, particularly in high-endemicity settings where latent TB infections (LTBI) contribute to ongoing transmission. Early identification and management of LTBI are crucial in limiting the spread of the disease. This study demonstrates the role of Interferon Gamma Release Assay (IGRA) as a screening tool for latent tuberculosis in high-burden region. Materials and Methods: This retrospective observational study assessed the detection of LTBI using the QuantiFERON-TB Gold Plus (QFT-Plus) test among 145 patients at the Department of Microbiology & Immunology, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh, from August 2023 to August 2024. The study included patients suspected of TB, those screened before immunosuppressive therapy, organ transplantation, or kidney dialysis. Participants were tested IGRA using QFT-Plus, which detects interferon-gamma (IFN-γ) released in response to Mycobacterium tuberculosis antigens. Results and Discussion: Among 145 patients tested for the QFT-Gold Plus test, 55.17% (n = 80) were positive for LTBI, with a substantial agreement between TB1 and TB2 responses (p Conclusion: The results highlight that QFT-Plus may be utilized as a useful diagnostic screening tool for latent TB in regions with a high disease burden, though challenges related to cost and infrastructure persist. With growing global efforts to eliminate tuberculosis, focused screening and treatment of LTBI in high-risk groups could play a vital role in reducing the progression of TB. The study underscores the importance of targeted screening for LTBI to reduce the progression to active TB, particularly in resource-limited settings. 展开更多
关键词 Latent Tuberculosis Infection (LTBI) Interferon-Gamma Release assay (IGRA) High Endemicity Regions LTBI Risk Factors
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基于Cytb基因的牦牛肉种来源鉴定与应用
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作者 何建文 党鹏举 +1 位作者 金丽娜 韩建林 《现代畜牧科技》 2025年第3期71-74,共4页
为建立牦牛、黄牛和水牛肉在基因水平快速、准确的鉴定方法,用于牦牛肉及制品的肉种来源鉴别研究。以牦牛、黄牛、水牛肉为研究对象,选用Cytb基因的一段保守区域对牦牛、黄牛和水牛序列进行限制性内切酶位点检测分析,发现3个特异性酶切... 为建立牦牛、黄牛和水牛肉在基因水平快速、准确的鉴定方法,用于牦牛肉及制品的肉种来源鉴别研究。以牦牛、黄牛、水牛肉为研究对象,选用Cytb基因的一段保守区域对牦牛、黄牛和水牛序列进行限制性内切酶位点检测分析,发现3个特异性酶切位点可用于牦牛肉及制品的肉种来源的鉴别。运用该方法对不同品牌的牦牛肉产品进行检测,再通过测序验证酶切结果的正确性及其黄牛肉、水牛肉假冒牦牛肉制品的参杂程度。结果显示,3个特异性酶切位点可用于混合牛肉产品的鉴别,牦牛肉制品的掺假现象较为严重。 展开更多
关键词 牦牛 pcr-rflp CYTB基因 物种鉴定
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马尾松叶际细菌多样性及其杀松材线虫活性 被引量:1
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作者 詹梦琳 何娇萍 +3 位作者 曾丽琼 何学友 林可竟 蔡学清 《生物安全学报(中英文)》 北大核心 2025年第1期86-95,共10页
【目的】分析马尾松叶际细菌种群多样性,挖掘其菌株资源;获得具有杀松材线虫活性的菌株,明确其分类地位,为松材线虫病的生物防治提供菌株资源。【方法】通过平板分离培养法从福建宁德古田、霞浦及三明尤溪的马尾松植株上分离叶际细菌,... 【目的】分析马尾松叶际细菌种群多样性,挖掘其菌株资源;获得具有杀松材线虫活性的菌株,明确其分类地位,为松材线虫病的生物防治提供菌株资源。【方法】通过平板分离培养法从福建宁德古田、霞浦及三明尤溪的马尾松植株上分离叶际细菌,通过菌落形态特征、16S rDNA PCR-RFLP对供试菌株进行分类;采用离心管培养法测定叶际细菌的杀松材线虫活性,通过生理生化特性测定、16S rDNA基因和gyrB基因序列分析明确拮抗菌株的分类地位。【结果】从福建宁德古田、霞浦及三明尤溪的马尾松植株上分离纯化获得141株马尾松叶际细菌,根据菌落形态特征将其分为9类,扩增获得的16S rDNA经HaeⅢ和MspⅠ酶切后分别有9和11种图谱类型,将2种酶切图谱组合后,在100%相似水平下,供试菌株共有19种图谱类型。以NB培养基或无菌水为对照,松材线虫的48 h校正死亡率为指标,经测定具有不同程度杀线虫效果的菌株有120株,占细菌总数的85.11%;以NB培养基为对照,松材线虫校正死亡率大于50%的菌株有11株,占筛选总数的7.80%,其中杀松材线虫效果最好的菌株为GT10,其48 h杀线虫率为92.56%;该菌株为革兰氏阳性菌,菌体杆状、周生鞭毛,可利用甘露醇和柠檬酸盐,硝酸还原反应和V-P试验呈阳性,系统发育树分析结果显示,菌株的16S rDNA基因和gyrB基因的序列均与蜡样芽胞杆菌模式菌株ATCC14579聚为一支,将菌株GT10鉴定为蜡样芽胞杆菌。【结论】马尾松叶际细菌具有丰富的多样性,其中蜡样芽胞杆菌菌株GT10具有较高的杀松材线虫活性,具有防治松材线虫病的潜力。 展开更多
关键词 马尾松 叶际细菌 pcr-rflp 松材线虫 蜡样芽胞杆菌
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IGF2基因PCR-RFLP多态性与脂肪沉积相关性状的关联分析 被引量:52
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作者 刘桂兰 蒋思文 +2 位作者 熊远著 郑嵘 屈彦纯 《Acta Genetica Sinica》 SCIE CAS CSCD 北大核心 2003年第12期1107-1112,共6页
胰岛素样生长因子 2在胎儿生长发育、肿瘤细胞增殖、肌肉生长等方面具有重要的调控作用 ,是影响猪瘦肉量的主要候选基因。采用PCR RFLP技术 ,分析了IGF2基因第 8内含子部分片段在猪资源家系群体中的NciⅠ酶切片段多态性分布。IGF2基因... 胰岛素样生长因子 2在胎儿生长发育、肿瘤细胞增殖、肌肉生长等方面具有重要的调控作用 ,是影响猪瘦肉量的主要候选基因。采用PCR RFLP技术 ,分析了IGF2基因第 8内含子部分片段在猪资源家系群体中的NciⅠ酶切片段多态性分布。IGF2基因该片段具有两个NciⅠ酶切位点 (切点位于第 8内含子 ,分别记录为位点A和位点B) ,两位点在资源家系中均具有多态性。IGF2基因B位点酶切未突变个体均比酶切突变的个体背膘薄 18 2 8% (P<0 0 1) ,肥肉率低 2 2 4 3% (P <0 0 1) ,瘦肉率高 8 71% (P <0 0 1) ,位点A具有相同的影响趋势。在该研究群体中 ,IGF2基因发挥作用的方式主要为加性效应 。 展开更多
关键词 IGF2基因 pcr-rflp 背膘厚
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我国主要地方绵羊品种mtDNA D-loop区PCR-RFLP研究 被引量:25
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作者 李祥龙 张增利 +3 位作者 巩元芳 刘铮铸 贾青 王立泽 《遗传》 CAS CSCD 北大核心 2006年第2期165-170,共6页
利用5种限制性内切酶(HinfⅠ,MspⅠ,Sau3AⅠ,XspⅠ,TaqⅠ),采用PCRRFLP技术研究了我国9个地方绵羊品种以及2个引入品种共计83只绵羊个体线粒体DNADloop区的多态性。结果表明,我国主要地方绵羊品种线粒体DNADloop区存在两种基本单体型,... 利用5种限制性内切酶(HinfⅠ,MspⅠ,Sau3AⅠ,XspⅠ,TaqⅠ),采用PCRRFLP技术研究了我国9个地方绵羊品种以及2个引入品种共计83只绵羊个体线粒体DNADloop区的多态性。结果表明,我国主要地方绵羊品种线粒体DNADloop区存在两种基本单体型,提示我国主要地方绵羊品种起源于两个母系祖先。线粒体DNADloop区多态度为0.0421%,说明我国地方绵羊品种线粒体DNA多态度较为贫乏。 展开更多
关键词 地方绵羊品种 线粒体DNA D-LOOP pcr-rflp
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细胞色素B基因PCR-RFLP鉴定阿胶原料 被引量:18
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作者 汪小龙 潘洁 +6 位作者 王师 万俊芬 包振民 解锡军 尤金花 解福生 师秀梅 《中国海洋大学学报(自然科学版)》 CAS CSCD 北大核心 2006年第4期645-648,共4页
用细胞色素B基因PCR-RFLP方法对阿胶原料进行鉴定。提取马、牛、驴3种动物干皮DNA,PCR扩增细胞色素B基因保守区域的359 bp片段,用限制性内切酶HinfⅠ和HaeⅢ酶切,采用琼脂糖凝胶电泳获得了DNA指纹图谱,根据获得的DNA指纹图谱判定样品所... 用细胞色素B基因PCR-RFLP方法对阿胶原料进行鉴定。提取马、牛、驴3种动物干皮DNA,PCR扩增细胞色素B基因保守区域的359 bp片段,用限制性内切酶HinfⅠ和HaeⅢ酶切,采用琼脂糖凝胶电泳获得了DNA指纹图谱,根据获得的DNA指纹图谱判定样品所属物种。该方法不仅简便,还可以100%准确鉴定阿胶原料皮样所属物种来源,适合作为常规技术应用于阿胶原料鉴定。 展开更多
关键词 物种鉴定 阿胶 CYT B pcr-rflp
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应用RAPD标记和细胞质基因组PCR-RFLP技术研究大花蕙兰的遗传多样性 被引量:16
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作者 甘娜 谭向红 +2 位作者 陈其兵 魏育明 郑有良 《园艺学报》 CAS CSCD 北大核心 2006年第2期349-355,共7页
利用RAPD、叶绿体和线粒体基因组PCRRFLP标记系统评价了大花蕙兰20个品种的遗传多样性。在50个RAPD引物分析中,有36个引物(72.0%)能揭示材料间的多态性,材料间遗传相似系数为0.503~0.765,平均0.598,根据遗传相似系数进行聚类分析表明,R... 利用RAPD、叶绿体和线粒体基因组PCRRFLP标记系统评价了大花蕙兰20个品种的遗传多样性。在50个RAPD引物分析中,有36个引物(72.0%)能揭示材料间的多态性,材料间遗传相似系数为0.503~0.765,平均0.598,根据遗传相似系数进行聚类分析表明,RAPD标记能将所有材料区分开。在7个叶绿体基因组(cpDNA)的PCRRFLP分析中,6个标记(87.5%)可扩增出1至多条清晰的谱带;扩增产物经7种限制性内切酶消化后,6个标记的19种引物/酶组合共检测到53条DNA片段,其中多态性片段有37条,占69.8%;材料间遗传相似系数变化范围为0.571~0.949,平均值为0.766。在8个线粒体基因组(mtDNA)的PCRRFLP标记分析中,只有3个(37.5%)标记能得到1条清晰的谱带;利用7种限制性内切酶对3个标记的扩增产物消化后,在10种标记/酶组合中,共检测到33条酶切片段,其中21条(63.6%)具有多态性;遗传相似系数为0.634~1.000,平均0.829。这些结果表明,RAPD标记揭示的大花蕙兰遗传多样性最高,其次为cpDNAPCRRFLP标记,而mtDNAPCRRFLP标记揭示的遗传多样性最低。 展开更多
关键词 大花蕙兰 RAPD pcr-rflp CPDNA MTDNA
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