LIU Xiao,LIN Cheng-xi,ZHANG Ping-ping,YANG Xiao-man,ZHAO Yong,LI Chun-feng,SUN Chong-yun,QIU Jing-fu,YANG Rui-fu,ZHOU Lei
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Objective To develop and evaluate an up-converting phosphor technology based on lateral flow assay (UPT-LF) for qualitative and quantitative detection of morphine (MOP) and methamphetamine (MET) in urine. Methods With up-converting phosphor nano-particles (UCP-NPs) as the biological tracer, two competitive mode-based LF strips, MOP-UPT-LF and MET-UPT-LF were developed for quantitative detection of MOP and MET in urine. The comprehensive performances of MOP-UPT-LF and MET-UPT-LF were evaluated systematically. In order to explore the detection limit of UPT-LF for trace analysis, MOP-UPT-LF strips were used to test standard samples with series of concentrations, and then the detection limit and ability of quantitative detection were determined. According to the standard of detection threshold for MOP and MET, the quantitative detection performances (including detection sensitivity and linear range) of MOP-UPT-LF and MET-UPT-LF were optimized and re-evaluated. For the evaluation of qualitative detection ability, the results of MOP-UPT-LF and MET-UPT-LF for on-site urine samples were compared with those of colloidal gold based LF (CG-LF), and LC-MS and GC-MS were used as the gold standard for the detection of MOP and MET, respectively. For the evaluation of quantitative detection ability, the results of MOP-UPT-LF and MET-UPT-LF for simulated positive urine samples were compared with those of LC-MS and GC-MS, respectively. Results For trace analysis, the detection limit of MOP-UPT-LF could reach 1ng/mL with a linear range from 1ng/mL to 5000ng/mL (r = -0.98172,P < 0.0005). For routine detection with the threshold of standard, the detection limit of MOP-UPT-LF was 50ng/mL with a linear range from 50ng/mL to 3000ng/mL (r = -0.98464,P < 0.0005). The detection limit of MET-UPT-LF was 100ng/mL with a linear range from 100ng/mL to 5000ng/mL (r = -0.99964, P < 0.0005). According to the detection of urine samples, the performance of qualitative and quantitative detection of MOP-UPT-LF and MET-UPT-LF could meet the need of the on-site rapid detection of MOP and MET in urine. The qualitative detection results of MOP-UPT-LF and MET-UPT-LF for on-site urine samples were consistent with those of colloidal gold and no false-positive and false-negative results observed. The ROC area of MOP-UPT-LF and MET-UPT-LF reached 1.000±0.000 (95%CI). For quantitative detection of simulated positive urine samples, the recovery rate was 77%~133% with the mean of 109% and CV of 21% for MOP-UPT-LF, for MET-UPT-LF the recovery rate was 80%~131% along with the mean of 112% and CV of 17%. After statistical analysis, there was no significant difference (P>0.05) between MOP-UPT-LF/MET-UPT-LF and LC-MS/GC-MS for quantitative detection of MOP and MET in urine. Conclusions With the novel optical nano-particle (UCP-NPs), the traditional LF assay was integrated with biosensor based on automated analysis and two kinds of UPT-LF strip were developed to meet the need of on-site qualitative and quantitative detection of MOP and MET. With GC-LF as the reference for qualitative detection and LC-MS/GC-MS for quantitative detection, the comprehensive performances (including detection limit, qualitative accuracy, and quantitative ability) were evaluated systematically. The good qualitative and quantitative detection performance of MOP-UPT-LF and MET-UPT-LF offers a new choice for on-site drug screening.