| Precision medical is a new disease prevention and treatment approach developed with the goal of individualized medical care,and the application of life analysis technology,big data and other interdisciplinary technologies.In addition,it is combined with patients'complete and accurate medical information and living environment.The key to accurate medical care includes the accurate use of drugs in clinic,fully considering the individual differences of patients,and preventing the side effects caused by drug overdose,or the insufficient dose cannot achieve the purpose of treatment.It is often impossible to achieve rapid,continuous and long-term monitoring of important information molecules or drugs in patients in the process of clinical drug detection.As a consequence,it is necessary to realize the accurate analysis and control of information molecules and drugs in patients,and to dynamically and quickly monitor their metabolism online.Moreover,it is very important to reduce the pain of patients,quickly respond to the sudden situation of patients(especially critically ill patients)and put forward emergency treatment plans in time.POCT(Point of Care Testing) has been developed rapidly as a new diagnostic mode.Taking vancomycin,a clinical"super antibiotic",as the target drug,this study optimizes the design of a variety of new fluorescent probes,and combines microdialysis sampling technology with microfluidic chip technology to achieve multi-point dynamic detection and continuous on-line monitoring of vancomycin without repeated collection of blood samples and complex sample pretreatment process.The problems of sample contamination and result delay time are eliminated.In addition,it has the advantages of short reaction time,high selectivity,high accuracy and good linear range.Moreover,what is more meaningful is that accurate pharmacokinetic information can be obtained timely and continuously.It can fundamentally change the existing empirical mode of drug administration in order to achieve the purpose of safe,reasonable,effective and economical drug use.In addition,it can provide a scientific and effective solution for the formulation of clinical high-precision drug use plan and individual drug efficacy and safety evaluation,and contribute to promote the practice and promotion of clinical precision medical treatment.The full text is divided into five parts,as follow.Chapter 1.OverviewIn this chapter,we mainly introduce the research background and significance of therapeutic drug monitoring(TDM)and antibiotics.In addition,it focuses on the physicochemical properties,toxic and side effects,application status and detection significance of vancomycin.Moreover,we also pay more attention to introduce of analytical method.Finally,the research purpose and significance of this paper,as well as the innovation spot and main research contents are pointed out.Chapter 2.Dual-emission fluorescence biosensing of vancomycin based on AIEgen peptide conjugates and aptamer-modified Au nanoclustersIn this study,a dual-emission fluorescence biosensor platform was developed for simple,selective,and sensitive determination of vancomycin(Van)based on a peptide conjugated with blue-emitting aggregation-induced emission luminogens(AIEgen)and aptamer-modified red-emitting gold nanoclusters(Au NCs-apt).The peptide and aptamer together recognized Van with high affinity,thus changing the fluorescence intensity at 470 nm and 650 nm,respectively.This platform displayed excellent linear correlation between the fluorescence response and a Van concentration ranging 0.01-100?g·mL-1,and the limit of detection(LOD) was 2.79 ng·mL-1.In addition to the ability to accurately distinguish Van from glycopeptide antibiotics,the newly developed biosensor allowed for naked-eye detection of 1?g·mL-1Van.These results and those of serum samples and microdialysate samples support the application of this newly developed method for Van monitoring and clinical diagnosis.It provides a new method and idea for POCT color test and chip test of clinical antibiotics in the future.Chapter 3.The development of a fluorescence biosensor for therapeutic drug monitoring of vancomycin using in vivo microdialysisClinical vancomycin treatment is administered over a prolonged period to achieve a sustained therapeutic effect.Accurate,rapid,and continuous Van detection is an unmet medical monitoring need that can provide data-based guidance for doctors to adjust the dosage and treatment plan in real-world settings.In this study,we created a Van-specific,fluorescent biosensor that was combined with a microdialysis sampling technique to develop a rapid,simple,accurate,and sensitive detection method,which was validated for Van in vivo.Subject-specific pharmacokinetics of Van was recorded in normal rabbits and rabbits with adenine-induced chronic renal failure(CRF),which indicated the feasibility of therapeutic drug monitoring in vivo.The area under the concentration–time curve(AUC0-last)was 10715 min·?g·mL-1(95%CI=8892 to 12538)in the normal rabbits,and 14822 and 19025 min·?g·mL-1 in two selected CRF rabbits.Furthermore,using pharmacokinetic dosing of Van in a rabbit study,we designed the dosage and drug administration interval to achieve a sustained therapeutic effect.The normal rabbits received three Van doses 20,5,and 5 mg·kg-1,administered at 0,500,and 900 min,respectively.The CRF rabbits received two Van doses 20 and 5 mg·kg-1,administered at 0 and 600 min,respectively.Thus,we established an effective method for the continuous monitoring of Van.This method facilitates the detection of Van in clinical treatment and provides the scientific basis for an effective approach to monitor blood drug levels during the clinical treatment of various diseases.Chapter 4.A novel “turn-off” fluorescent probe for the construction of online monitoring platform for vancomycinIn this work,we designed and synthesized a"turn-off"fluorescent probe with double fluorescent groups and double recognition groups for the detection of vancomycin,and combined it with a 375 nm laser-induced fluorescence laser for verification on the online detection platform.We explored the influence of different factors on the construction of online detection system.The influence of path diameter,path length,solution flow rate,the concentration of fluorescent probe and the effect of mixing on online fluorescence signal were detected.Compared to commercial micro-reactor,we designed the 3D microchip to have a smaller volume(about a thumbnail),smaller liquid holdup(about 10?L),good mixing effect and fast signal response time (within 1 min).The development of this work provides a scientific model for the construction of an online detection system.Chapter 5.Online POCT detection system for monitoring of vancomycin based on“turn-on”fluorescent probe with an aggregation-induced emission characteristicsThis work is based on the construction of vancomycin online drug detection platform in the previous chapter.We optimized the design and synthesis of an aggregation-induced fluorescence molecular probe for the stable 405 nm excitation-induced fluorescence source for the detection of vancomycin.The fluorescent probe is a"turn-on"fluorescent probe,which has a higher signal-to-noise ratio and sensitivity than the fluorescent probe designed in previous work.In addition,it has high resistance to photobleaching under strong excitation light.On this basis,we constructed in vivo microdialysis sampling-fluorescence sensor online detection platform.And the microchip was used as the reaction and detection unit.A laser with a stable excitation source was selected for molecular recognition and sensing analysis.The platform had been successfully applied to monitor the metabolism of vancomycin in animals.Different from the multi-point dynamic sampling detection,an online real-time monitoring system for the concentration of antibiotics in blood was established.The system can realize online,continuous detection,accurate diagnosis and scientific drug use.In addition,it can provide a scientific and effective solution for the formulation of clinical high-precision drug use plan and individual drug efficacy and safety evaluation. |
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