| Heart failure(HF)is a complicated clinical syndrome with high mortality and readmission rate,and it is a common critical disease in children,which may endanger the life of the child without timely diagnosis and treatment.The monitoring of biomarkers related to HF is of significant importance to the early diagnosis and timely intervention of HF.Brain natriuretic peptide(BNP)has been an internationally recognized biomarker of HF since its secretion is significantly increased in the blood of HF patients owing to intravascular volume expansion and pressure overload in their hearts.A variety of different methods have been developed for BNP assay,including chemiluminescence immunoassay(CLIA),immunochromatography assay(ICA),enzyme?linked immunosorbent assay(ELISA),and so on.However,these methods suffer from some limitations such as poor sensitivity and relatively weak anti?interference ability.Thus,two fluorescence aptasensor based on nano graphene oxide and cobalt oxyhydroxide were fabricated for the detection of BNP.The details of the researches are as follows:Part Ⅰ Graphene oxide?based fluorescence aptasensor for the ultrasensitive detection of brain natriuretic peptideIn order to achieve the early diagnosis of pediatric HF,we developed a fluorescence resonance energy transfer(FRET)platform for the detection of BNP,a biomarker of HF.Graphene oxide(GO)was selected as the FRET receptor in view of its advantages including commercial availability,low cost and chemical stability,and dye?modified aptamer was used as the energy donor of FRET as well as in charge of the specific recognition of BNP.Based on the ON(strong emission)and OFF(quenching)states of FRET in the presence and absence of BNP,respectively,specific detection of BNP was achieved in the range of 0.074?0.56 pg/mL with a limit of detection as low as 0.045 pg/mL.This FRET platform was further applied to detect BNP in 45 blood samples of suspected pediatric HF to demonstrate its practicability in clinical diagnosis.Compared with the commonly used Siemens method(CLIA)in hospital,our approach is more accurate and specific for HF diagnosis with areas under the receiver operating characteristic curves of 0.869(95% CI: 0.733?1.00,P < 0.05)vs 0.850(95% CI: 0.703?0.997,P < 0.05)and specificity of 68.8% vs 65.6%.This platform is promising in early diagnosis of HF through ultrasensitive and specific detection of BNP.Part Ⅱ A fluorescence resonance energy transfer platform based on cobalt oxyhydroxide nanorings and aptamer for the detection of brain natriuretic peptideA FRET platform for the sensitive and selective detection of BNP was fabricated using cobalt oxyhydroxide nanorings(CoOOH NRs)as energy receptor and FAM?labeled aptamer as energy receptor and recognition element.In the absence of BNP,CoOOH NRs can adsorb the aptamer and thereby quenching the fluorescence of FAM?aptamer through FRET.However,in the presence of target protein BNP,the aptamer can combine with BNP through the specific molecular recognition,leading to the conformation change of the aptamer and thus hindering its adsorption onto the CoOOH NRs.In this case,the fluorescence can be restored.On the basis of such a mechanism,the ultrasensitive and selective detection of BNP was achieved in the range of 0.030?0.80 pg/mL with a limit of detection as low as 0.030 pg/mL.By detecting as many as 57 blood samples from suspected pediatric HF patients,we have demonstrated that our method has a close correlation(r = 0.92,P = 0.00)to that of Siemens method.With the aid of ROC curve analysis,these two methods(our method vs Siemens method)exhibited good diagnosis performance with the areas under the curves of 0.774(95% CI: 0.573?0.974,P < 0.05)vs 0.829(95% CI: 0.686?0.972,P < 0.05).However,the specificity(84.6% vs 61.5%)and positive predictive value(40.0% vs 20.0%)of diagnosis have improved in our method.The accurate detection of BNP in complicated blood and therefore the early diagnosis of pediatric HF can be achieved by this rapid,cost?effective,ultrasensitive and selective method. |
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