Phytohemagglutinin(PHA)is a seed storage protein and a type of lectin originally discovered in the common bean(Phaseolus vulgaris)for its blood-agglutinating effect.Due to its interactions with gut epithelia and diges...Phytohemagglutinin(PHA)is a seed storage protein and a type of lectin originally discovered in the common bean(Phaseolus vulgaris)for its blood-agglutinating effect.Due to its interactions with gut epithelia and digestive enzymes and its potential to trigger allergic reactions,PHA can lead to various symptoms in the human body.As a result,it has been regarded as a signifcant antinutritional factor in beans and other legumes.While several published works have summarized its structural,biochemical,and toxicological features,there is a scarcity of literature that reviews the detection,quantifcation,and reduction of PHA in beans,which is fundamental for the development of safer bean varieties.In this review,we present a comprehensive analysis of traditional and innovative bio-sensing methods for measuring PHA,including the recently available ultrapure liquid chromatography–tandem mass spectrometry and emerging aptamer sensor-based techniques,while discussing their respective advantages and disadvantages.We also revisit existing studies dedicated to creating PHA-depleted common bean varieties and explore the potential for reducing PHA content in beans without compromising their resistance to biotic stress.Additionally,we offer insights into the potential for controlling PHA content using the latest biotechnologies and breeding strategies.Overall,this review compiles rare and valuable information from studies that solely focuses on detection and depletion of PHA to shed light on and apply technological advancements in addressing potential food safety risks associated with the consumption of common beans.展开更多
目的:建立一种快速、高效、可视化的细菌多黏菌素耐药基因mcr-1检测方法,为其基层检测的展开提供依据和便利。方法:利用重组酶聚合酶扩增结合胶体金侧向流试纸条技术(Recombinase polymerase amplification combined with a lateral flo...目的:建立一种快速、高效、可视化的细菌多黏菌素耐药基因mcr-1检测方法,为其基层检测的展开提供依据和便利。方法:利用重组酶聚合酶扩增结合胶体金侧向流试纸条技术(Recombinase polymerase amplification combined with a lateral flow dipstick,RPA-LFD),辅以手持式胶体金读数仪;根据mcr-1基因保守序列设计合成一对特异性RPA引物,通过对反应条件和体系的优化,以及特异性试验、灵敏度试验、模拟食样试验和实际样品试验,成功建立了可视化定量检测细菌多黏菌素耐药基因mcr-1的RPA-LFD方法。结果:在引物浓度400 nmol/L,引物比例1:1时,该方法最佳反应条件为Mg^(2+)浓度14.0 mmol/L,反应温度37℃,反应时间20 min;灵敏度好,标准曲线方程为y=0.117x+0.051,定量限为10^(1)~108 copies/μL,检出限为10^(1)copies/μL,比PCR法低一个数量级且模拟样品检出结果与PCR法一致。利用建立的RPA-LFD法对猪肉样品、鸡肉样品、生猪养殖场环境样品、肉鸡养殖场环境样品、大肠杆菌分离株和弯曲肠杆菌分离株各15份中多黏菌素耐药基因mcr-1携带情况进行分析;RPA-LFD法与常规PCR法阳性样本检出率一致,共检出9份mcr-1基因阳性样品。RPA-LFD定量分析显示,阳性样品中mcr-1基因浓度在4.5×10^(2)~8.6×10^(4)copies/μL之间。结论:本研究建立的细菌多黏菌素耐药基因mcr-1的RPA-LFD检测法特异性强、灵敏性高、操作简单,可广泛应用于基层检验。展开更多
基金the National Natural Science Foundation of China(Nos.32201254 and 32302545)the Basic Public Welfare Research Program of Zhejiang Province(No.LTGN23C020003)the State Key Laboratory for Managing Biotic and Chemical Treats to the Quality and Safety of Agro-products(No.2021DG700024-KF202403),China.
文摘Phytohemagglutinin(PHA)is a seed storage protein and a type of lectin originally discovered in the common bean(Phaseolus vulgaris)for its blood-agglutinating effect.Due to its interactions with gut epithelia and digestive enzymes and its potential to trigger allergic reactions,PHA can lead to various symptoms in the human body.As a result,it has been regarded as a signifcant antinutritional factor in beans and other legumes.While several published works have summarized its structural,biochemical,and toxicological features,there is a scarcity of literature that reviews the detection,quantifcation,and reduction of PHA in beans,which is fundamental for the development of safer bean varieties.In this review,we present a comprehensive analysis of traditional and innovative bio-sensing methods for measuring PHA,including the recently available ultrapure liquid chromatography–tandem mass spectrometry and emerging aptamer sensor-based techniques,while discussing their respective advantages and disadvantages.We also revisit existing studies dedicated to creating PHA-depleted common bean varieties and explore the potential for reducing PHA content in beans without compromising their resistance to biotic stress.Additionally,we offer insights into the potential for controlling PHA content using the latest biotechnologies and breeding strategies.Overall,this review compiles rare and valuable information from studies that solely focuses on detection and depletion of PHA to shed light on and apply technological advancements in addressing potential food safety risks associated with the consumption of common beans.
