Droplet digital PCR(ddPCR),as the third-generation PCR technology,demonstrates significant advantages in the etiological diagnosis of infectious diseases due to its absolute quantification,ultra-high sensitivity,and m...Droplet digital PCR(ddPCR),as the third-generation PCR technology,demonstrates significant advantages in the etiological diagnosis of infectious diseases due to its absolute quantification,ultra-high sensitivity,and multiplex detection capabilities.This article reports a case of a patient with fever of unknown origin,where ddPCR rapidly confirmed a drug-resistant bacterial infection and dynamically monitored treatment efficacy.Combining literature evidence,this paper systematically elaborates on the technical principles,clinical performance,and practical value of ddPCR in febrile patients.展开更多
Brucellosis is a global public health issue that severely affects human health,Brucella melitensis is currently the predominant species in China.Brucella spondylitis is the primary cause of the debilitating and disabl...Brucellosis is a global public health issue that severely affects human health,Brucella melitensis is currently the predominant species in China.Brucella spondylitis is the primary cause of the debilitating and disabling complications[1].The lumbar vertebra was the most commonly affected site,followed by the thoracic,cervical,thoracolumbar,and lumbosacral segments,and back pain,fever,sweating,and fatigue were the most common symptoms[2].However,the diagnosis of Brucella spondylitis is challenging owing to its wide spectrum of clinical presentations,cross-reactions with other bacteria,and low strain isolation rate.Therefore,a timely and accurate diagnosis of spinal brucellosis is crucial for implementing an effective therapeutic plan and improving treatment outcomes.Droplet digital polymerase chain reaction(ddPCR)is widely used for low-abundance nucleic acid detection and is useful for diagnosing infectious diseases[3].Therefore,this study aimed to evaluate the ddPCR approach for the diagnosis of brucellosis with spondylitis to improve its clinical diagnostic capacity.展开更多
目的建立炭疽芽孢杆菌的微滴式数字聚合酶链式反应(Droplet digital PCR,ddPCR)方法对炭疽芽孢杆菌实验室活动污染的定量评估提供技术支持。方法以炭疽芽孢杆菌pXO1质粒编码保护性抗原pagA基因为靶序列,优化微滴数字PCR方法的反应条件,...目的建立炭疽芽孢杆菌的微滴式数字聚合酶链式反应(Droplet digital PCR,ddPCR)方法对炭疽芽孢杆菌实验室活动污染的定量评估提供技术支持。方法以炭疽芽孢杆菌pXO1质粒编码保护性抗原pagA基因为靶序列,优化微滴数字PCR方法的反应条件,建立实验室微环境中炭疽芽孢杆菌核酸定量方法;对比微滴式数字PCR方法和平板计数法的定量评估效果,分析ddPCR的灵敏性、特异性和重复性。结果建立的ddPCR方法最佳引物和探针终浓度分别为900nmol·L^(-1)和250nmol·L^(-1),最佳退火温度为60℃,最佳升降温速度为1℃/s,本方法的最低检测下限为1.12copies·μL^(-1),未发现与常见疫病存在交叉反应,重复性试验的变异系数小于5%。结论本研究中建立的炭疽芽孢杆菌的微滴数字PCR方法敏感性高、特异性强、重复性好,为疫情监测、流行病学调查和实验室污染微环境检测提供重要技术。展开更多
文摘Droplet digital PCR(ddPCR),as the third-generation PCR technology,demonstrates significant advantages in the etiological diagnosis of infectious diseases due to its absolute quantification,ultra-high sensitivity,and multiplex detection capabilities.This article reports a case of a patient with fever of unknown origin,where ddPCR rapidly confirmed a drug-resistant bacterial infection and dynamically monitored treatment efficacy.Combining literature evidence,this paper systematically elaborates on the technical principles,clinical performance,and practical value of ddPCR in febrile patients.
基金supported by the Key Research and Development Projects of the Ningxia Hui Autonomous Region(Grant no.2022BEG03161)。
文摘Brucellosis is a global public health issue that severely affects human health,Brucella melitensis is currently the predominant species in China.Brucella spondylitis is the primary cause of the debilitating and disabling complications[1].The lumbar vertebra was the most commonly affected site,followed by the thoracic,cervical,thoracolumbar,and lumbosacral segments,and back pain,fever,sweating,and fatigue were the most common symptoms[2].However,the diagnosis of Brucella spondylitis is challenging owing to its wide spectrum of clinical presentations,cross-reactions with other bacteria,and low strain isolation rate.Therefore,a timely and accurate diagnosis of spinal brucellosis is crucial for implementing an effective therapeutic plan and improving treatment outcomes.Droplet digital polymerase chain reaction(ddPCR)is widely used for low-abundance nucleic acid detection and is useful for diagnosing infectious diseases[3].Therefore,this study aimed to evaluate the ddPCR approach for the diagnosis of brucellosis with spondylitis to improve its clinical diagnostic capacity.
文摘目的建立炭疽芽孢杆菌的微滴式数字聚合酶链式反应(Droplet digital PCR,ddPCR)方法对炭疽芽孢杆菌实验室活动污染的定量评估提供技术支持。方法以炭疽芽孢杆菌pXO1质粒编码保护性抗原pagA基因为靶序列,优化微滴数字PCR方法的反应条件,建立实验室微环境中炭疽芽孢杆菌核酸定量方法;对比微滴式数字PCR方法和平板计数法的定量评估效果,分析ddPCR的灵敏性、特异性和重复性。结果建立的ddPCR方法最佳引物和探针终浓度分别为900nmol·L^(-1)和250nmol·L^(-1),最佳退火温度为60℃,最佳升降温速度为1℃/s,本方法的最低检测下限为1.12copies·μL^(-1),未发现与常见疫病存在交叉反应,重复性试验的变异系数小于5%。结论本研究中建立的炭疽芽孢杆菌的微滴数字PCR方法敏感性高、特异性强、重复性好,为疫情监测、流行病学调查和实验室污染微环境检测提供重要技术。
