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Development and evaluation of an up-converting phosphor technology-based lateral flow assay for rapid detection of Francisella tularensis.

Hua F, Zhang P, Zhang F, Zhao Y, Li C, Sun C, Wang X, Yang R, Wang C, Yu A, Zhou L - Sci Rep (2015)

Bottom Line: Except Shigella dysenteriae, UPT-LF showed excellent specificity to four strains that are also potential biowarfare/bioterrorism agents and 13 food-borne pathogenic strains.For practical utilization, the tolerance limits for seven powders and eight viscera were determined, and operation errors of liquid measurement demonstrated a minor influence on the strip.Ftu-UPT-LF is a candidate POCT method because of its excellent sensitivity, specificity, and stability in complex samples, as well as low operation error.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, P.R. China.

ABSTRACT
Francisella tularensis is a potential biowarfare/bioterrorism agent and zoonotic pathogen that causes tularemia; thus, surveillance of F. tularensis and first-level emergency response using point-of-care testing (POCT) are essential. The UPT-LF POCT assay was established to quantitatively detect F. tularensis within 15 min, and the sensitivity of the assay was 10(4) CFU · mL(-1) (100 CFU/test). The linear quantitative range covered five orders of magnitude, and the coefficients of variation were less than 10%. Except Shigella dysenteriae, UPT-LF showed excellent specificity to four strains that are also potential biowarfare/bioterrorism agents and 13 food-borne pathogenic strains. Samples with pH 2-13, high ion strengths (≥ 2 mol · L(-1) solution of KCl and NaCl), high viscosities (≤ 50 mg · mL(-1) PEG20000 or ≥ 20% glycerol), and high concentrations of biomacromolecules (≥ 400 mg · mL(-1) bovine serum albumin or ≥ 80 mg · mL(-1) casein) showed little influence on the assay. For practical utilization, the tolerance limits for seven powders and eight viscera were determined, and operation errors of liquid measurement demonstrated a minor influence on the strip. Ftu-UPT-LF is a candidate POCT method because of its excellent sensitivity, specificity, and stability in complex samples, as well as low operation error.

No MeSH data available.


Related in: MedlinePlus

T/C ratios of Ftu-UPT-LF under the influence of (a) pH, (b) ionstrength, (c) viscosity, and (d) biological matrices. The linegraphs represent the T/C ratios of all samples with F. tularensisranging from104 CFU · mL−1to108 CFU · mL−1,whereas the bar graphs show the results of negative and weak positivesamples in detail. Ftu-UPT-LF maintained sensitivity and specificity (*) atcertain concentrations of the single-factor interference.
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f3: T/C ratios of Ftu-UPT-LF under the influence of (a) pH, (b) ionstrength, (c) viscosity, and (d) biological matrices. The linegraphs represent the T/C ratios of all samples with F. tularensisranging from104 CFU · mL−1to108 CFU · mL−1,whereas the bar graphs show the results of negative and weak positivesamples in detail. Ftu-UPT-LF maintained sensitivity and specificity (*) atcertain concentrations of the single-factor interference.

Mentions: Serial concentrations of HCl and NaOH, the mixture of KCl and NaCl, PEG20000 andglycerol, and BSA and casein were used to assess the single-factor theoreticaltolerance of Ftu-UPT-LF to pH, ion strength, viscosity, and biological matrices,respectively. Under the influence of various chemical and biological agents, theT/C ratios of all samples with F. tularensis ranging from104 CFU · mL−1to108 CFU · mL−1showed favorable consistency with that of the control (line graph in Fig. 3). The error in quantitative analysis of Ftu-UPT-LFwas less than one order of magnitude, indicating the accuracy of quantitativedetection.


Development and evaluation of an up-converting phosphor technology-based lateral flow assay for rapid detection of Francisella tularensis.

Hua F, Zhang P, Zhang F, Zhao Y, Li C, Sun C, Wang X, Yang R, Wang C, Yu A, Zhou L - Sci Rep (2015)

T/C ratios of Ftu-UPT-LF under the influence of (a) pH, (b) ionstrength, (c) viscosity, and (d) biological matrices. The linegraphs represent the T/C ratios of all samples with F. tularensisranging from104 CFU · mL−1to108 CFU · mL−1,whereas the bar graphs show the results of negative and weak positivesamples in detail. Ftu-UPT-LF maintained sensitivity and specificity (*) atcertain concentrations of the single-factor interference.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4660431&req=5

f3: T/C ratios of Ftu-UPT-LF under the influence of (a) pH, (b) ionstrength, (c) viscosity, and (d) biological matrices. The linegraphs represent the T/C ratios of all samples with F. tularensisranging from104 CFU · mL−1to108 CFU · mL−1,whereas the bar graphs show the results of negative and weak positivesamples in detail. Ftu-UPT-LF maintained sensitivity and specificity (*) atcertain concentrations of the single-factor interference.
Mentions: Serial concentrations of HCl and NaOH, the mixture of KCl and NaCl, PEG20000 andglycerol, and BSA and casein were used to assess the single-factor theoreticaltolerance of Ftu-UPT-LF to pH, ion strength, viscosity, and biological matrices,respectively. Under the influence of various chemical and biological agents, theT/C ratios of all samples with F. tularensis ranging from104 CFU · mL−1to108 CFU · mL−1showed favorable consistency with that of the control (line graph in Fig. 3). The error in quantitative analysis of Ftu-UPT-LFwas less than one order of magnitude, indicating the accuracy of quantitativedetection.

Bottom Line: Except Shigella dysenteriae, UPT-LF showed excellent specificity to four strains that are also potential biowarfare/bioterrorism agents and 13 food-borne pathogenic strains.For practical utilization, the tolerance limits for seven powders and eight viscera were determined, and operation errors of liquid measurement demonstrated a minor influence on the strip.Ftu-UPT-LF is a candidate POCT method because of its excellent sensitivity, specificity, and stability in complex samples, as well as low operation error.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, P.R. China.

ABSTRACT
Francisella tularensis is a potential biowarfare/bioterrorism agent and zoonotic pathogen that causes tularemia; thus, surveillance of F. tularensis and first-level emergency response using point-of-care testing (POCT) are essential. The UPT-LF POCT assay was established to quantitatively detect F. tularensis within 15 min, and the sensitivity of the assay was 10(4) CFU · mL(-1) (100 CFU/test). The linear quantitative range covered five orders of magnitude, and the coefficients of variation were less than 10%. Except Shigella dysenteriae, UPT-LF showed excellent specificity to four strains that are also potential biowarfare/bioterrorism agents and 13 food-borne pathogenic strains. Samples with pH 2-13, high ion strengths (≥ 2 mol · L(-1) solution of KCl and NaCl), high viscosities (≤ 50 mg · mL(-1) PEG20000 or ≥ 20% glycerol), and high concentrations of biomacromolecules (≥ 400 mg · mL(-1) bovine serum albumin or ≥ 80 mg · mL(-1) casein) showed little influence on the assay. For practical utilization, the tolerance limits for seven powders and eight viscera were determined, and operation errors of liquid measurement demonstrated a minor influence on the strip. Ftu-UPT-LF is a candidate POCT method because of its excellent sensitivity, specificity, and stability in complex samples, as well as low operation error.

No MeSH data available.


Related in: MedlinePlus