Development and evaluation of an up-converting phosphor technology-based lateral flow assay for the rapid, simultaneous detection of Vibrio cholerae serogroups O1 and O139

Abstract

Vibrio cholerae serogroups O1 and O139 are etiological agents of cholera, a serious and acute diarrheal disease, and rapid detection of V. cholerae is a key method for preventing and controlling cholera epidemics. Here, a point of care testing (POCT) method called Vch-UPT-LF, which is an up-converting phosphor technology-based lateral flow (UPT-LF) assay with a dual-target detection mode, was developed to detect V. cholerae O1 and O139 simultaneously from one sample loading. Although applying an independent reaction pair made both detection results for the two Vch-UPT-LF detection channels more stable, the sensitivity slightly declined from 104 to 105 colony-forming units (CFU) mL−1 compared with that of the single-target assay, while the quantification ranges covering four orders of magnitude were maintained. The strip showed excellent specificity for seven Vibrio species that are highly related genetically, and nine food-borne species whose transmission routes are similar to those of V. cholerae. The legitimate arrangement of the two adjacent test lines lessened the mutual impact of the quantitation results between the two targets, and the quantification values did not differ by more than one order of magnitude when the samples contained high concentrations of both V. cholerae O1 and O139. Under pre-incubation conditions, 1×101 CFU mL−1 of V. cholerae O1 or O139 could be detected in fewer than 7 h, while the Vch-UPT-LF assay exhibited sensitivity as high as a real-time fluorescent polymerase chain reaction with fewer false-positive results. Therefore, successful development of Vch-UPT-LF as a dual-target assay for quantitative detection makes this assay a good candidate POCT method for the detection and surveillance of epidemic cholera.

Calibration of an Upconverting Phosphor-Based Quantitative Immunochromatographic Assay for Detecting Yersinia pestis, Brucella spp., and Bacillus anthracis Spores

“…(Zhao et al, 2016). Our UPT-LF assays showed robust performance using many clinical and environmental samples such as biochemical reagents and various types of powders and viscera samples (Zhang P. et al, 2014;Hua et al, 2015a,b), as well as field water samples (Hao et al, 2017). After the samples are loaded onto the strip, upconverting phosphor (UCP) particles that have first been combined with monoclonal antibodies (mAbs) against bacteria can capture the corresponding bacteria and are then captured by the other antibodies against bacteria fixed onto the test band (T band), while the remaining UCP-mAbs complexes are captured by goat anti-mouse IgG on the control band (C band).…”

“…UPT-LF assays for the detection of pathogenic bacteria have been developed and evaluated in our laboratory, and include assays to detect Y. pestis (Yan et al, 2006), B. anthracis spores (Li et al, 2006), Brucella spp. (Qu et al, 2009), Vibrio cholerae (Hao et al, 2017), Burkholderia pseudomallei (Hua et al, 2015a;Liang et al, 2017), Francisella tularensis (Hua et al, 2015b), Escherichia coli O157:H7 (Wang et al, 2007), Vibrio parahaemolyticus, and seven Salmonella spp. (Zhao et al, 2016).…”

Multiplex Lateral Flow Immunoassay: An Overview of Strategies Towards High-Throughput Point-of-Need Testing

“…Multiplex LFIA realized through the spatial separation of lines in one strip has been combined also with the exploitation of innovative labels, such as up-converting phosphor reporter particles [36][37], magnetic nanobeads [38][39], enzymes triggering chemiluminescence [40] and near-infrared fluorescence label [41]. In the last paper, the detection of three classes of antimicrobial agents in a single run was achieved by combining the use of three broad-specific antibodies and the spatial separation of three lines in one strip.…”

Dual Detection of Procalcitonin and C-reactive Protein with an Up-converting Nanoparticle Based Lateral Flow Assay

“…19 The upconverting phosphor technology-based lateral flow assay (UPT-LFA) has been widely used in immunoassays, and it is suitable for multiple detection as well. 20 Multiple detection is much more difficult than is single detection, not only for the improvement demand of instruments, but also for cross interference caused by non-specific interactions and spectral interference. Many efforts have been put into multiple detection, including pathogens, microorganisms, medicinal metabolites, and food and environmental monitoring.…”

“…Many efforts have been put into multiple detection, including pathogens, microorganisms, medicinal metabolites, and food and environmental monitoring. [20][21][22][23] For example, Liang et al 24 synthesized single UCNPs for the multiple detection of bacteria with green and red emissions, which may have problems with non-specific adsorption. Another difficulty of the simultaneous detection of PCT and CRP is the substantial differences in concentration levels (ng/mL for PCT and μg/mL for CRP).…”