Comparison of Hand-Held Test Kits, Immunofluorescence Microscopy, Enzyme-Linked Immunosorbent Assay, and Flow Cytometric Analysis for Rapid Presumptive Identification of
            <i>Yersinia pestis</i>

Tomaso, Herbert; Thullier, Philippe; Seibold, Erik; Guglielmo, V.; Buckendahl, A.; Rahalison, Lila; Neubauer, Heinrich; Scholz, Holger C.; Splettstoesser, Wolf D.

doi:10.1128/jcm.00458-07

Cited by 31 publications

(39 citation statements)

References 10 publications

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“…Commercially available diagnostics such as the Plague BioThreat Alert test strips are based on the detection of the fraction 1 capsular antigen. The sensitivity limits of detection are between 6 ϫ 10 3 and 7 ϫ 10 3 CFU/ml, which is similar to the detection sensitivity described in this report (33). However, both of these sensitivity limits pale in comparison with that of real-time PCR, which can detect between 0.01 and 1 pg of extracted DNA; assuming a 100% extraction efficiency, this translates to approximately 1 CFU (24,32).…”

Section: Discussionsupporting

confidence: 51%

“…Y. pestis A1122 is an exempt select-agent strain (26) which lacks the pgm locus and the pCD1 plasmid, both of which encode critical virulence factors. Attenuated strains are commonly used for research purposes in order to demonstrate diagnostic detection feasibility (21,31,33). Although we used an attenuated strain in this study, there is strong evidence to indicate that the results obtained will translate to fully virulent Y. pestis strains.…”

Section: Discussionmentioning

confidence: 99%

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Diagnostic Bioluminescent Phage for Detection of Yersinia pestis

2009

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Yersinia pestis is the etiological agent of the plague. Because of the disease's inherent communicability, rapid clinical course, and high mortality, it is critical that an outbreak, whether it is natural or deliberate, be detected and diagnosed quickly. The objective of this research was to generate a recombinant luxAB ("light")-tagged reporter phage that can detect Y. pestis by rapidly and specifically conferring a bioluminescent signal response to these cells. The bacterial luxAB reporter genes were integrated into a noncoding region of the CDC plague-diagnostic phage A1122 by homologous recombination. The identity and fitness of the recombinant phage were assessed through PCR analysis and lysis assays and functionally verified by the ability to transduce a bioluminescent signal to recipient cells. The reporter phage conferred a bioluminescent phenotype to Y. pestis within 12 min of infection at 28°C. The signal response time and signal strength were dependent on the number of cells present. A positive signal was obtained from 10 2 cells within 60 min. A signal response was not detectable with Escherichia coli, although a weak signal (100-fold lower than that with Y. pestis) was obtained with 1 (of 10) Yersinia enterocolitica strains and 2 (of 10) Yersinia pseudotuberculosis strains at the restrictive temperature. Importantly, serum did not prevent the ability of the reporter phage to infect Y. pestis, nor did it significantly quench the resulting bioluminescent signal. Collectively, the results indicate that the reporter phage displays promise for the rapid and specific diagnostic detection of cultivated Y. pestis isolates or infected clinical specimens.

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Section: Discussionsupporting

confidence: 51%

Section: Discussionmentioning

confidence: 99%

Diagnostic Bioluminescent Phage for Detection of Yersinia pestis

2009

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“…For example, the F1 antigen could be detected at a concentration of 0.5 ng/ml, while 3 ϫ 10 3 CFU/ml of plague bacilli gave a positive reaction (9,10,13,46). Specifically, a recent study with the same Plague BioThreat Alert test strips from Tetracore reported a detection limit of 37 ng/ml for the purified F1 antigen and 6 ϫ 10 3 CFU/ml for the bacteria (46).…”

Section: Vol 49 2011 Rapid Identification Of Capsular Y Pestis Strmentioning

confidence: 99%

“…Our studies are the first to use these kits with a highly virulent CO92 Y. pestis strain and its authentic F1 mutant. The presence of the F1 antigen in plague patient sera has been reported in several studies and ranged from as low as 4 ng/ml to as high as 50,000 ng/ml (8,46,51). However, antigenemia was not present in all patients with acute plague, and only 20% of their sera were positive for the Fl antigen (51).…”

Section: Vol 49 2011 Rapid Identification Of Capsular Y Pestis Strmentioning

confidence: 99%

Characterization of an F1 Deletion Mutant of Yersinia pestis CO92, Pathogenic Role of F1 Antigen in Bubonic and Pneumonic Plague, and Evaluation of Sensitivity and Specificity of F1 Antigen Capture-Based Dipsticks

et al. 2011

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We evaluated two commercial F1 antigen capture-based immunochromatographic dipsticks, Yersinia Pestis (F1) Smart II and Plague BioThreat Alert test strips, in detecting plague bacilli by using whole-blood samples from mice experimentally infected with Yersinia pestis CO92. To assess the specificities of these dipsticks, an in-frame F1-deficient mutant of CO92 (⌬caf) was generated by homologous recombination and used as a negative control. Based on genetic, antigenic/immunologic, and electron microscopic analyses, the ⌬caf mutant was devoid of a capsule. The growth rate of the ⌬caf mutant generally was similar to that of the wild-type (WT) bacterium at both 26 and 37°C, although the mutant's growth dropped slightly during the late phase at 37°C. The ⌬caf mutant was as virulent as WT CO92 in the pneumonic plague mouse model; however, it was attenuated in developing bubonic plague. Both dipsticks had similar sensitivities, requiring a minimum of 0.5 g/ml of purified F1 antigen or 1 ؋ 10 5 to 5 ؋ 10 5 CFU/ml of WT CO92 for positive results, while the blood samples were negative for up to 1 ؋ 10 8 CFU/ml of the ⌬caf mutant. Our studies demonstrated the diagnostic potential of two plague dipsticks in detecting capsular-positive strains of Y. pestis in bubonic and pneumonic plague.

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“…BioThreat Alert Ò Assays have previously been evaluated for the detection of other biothreat agents, including orthopoxviruses, 50 ricin, 33 abrin, 34 and Yersinia pestis. 51 Limited evaluations have also been conducted with LFAs for the detection of Francisella tularensis (unpublished data), botulinum neurotoxins, 52 and staphylococcal enterotoxins. 53 In an earlier study, King et al 54 detected 10 5 spores/mL of B. anthracis Pasteur strain using the Tetracore BTA LFA.…”

Section: Discussionmentioning

confidence: 99%

Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification ofBacillus anthracisSpores in Suspicious White Powders and Environmental Samples

et al. 2016

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We conducted a comprehensive, multiphase laboratory evaluation of the Anthrax BioThreat Alert Ò test strip, a lateral flow immunoassay (LFA) for the rapid detection of Bacillus anthracis spores. The study, conducted at 2 sites, evaluated this assay for the detection of spores from the Ames and Sterne strains of B. anthracis, as well as those from an additional 22 strains. Phylogenetic near neighbors, environmental background organisms, white powders, and environmental samples were also tested. The Anthrax LFA demonstrated a limit of detection of about 10 6 spores/mL (ca. 1.5 · 10 5 spores/assay). In this study, overall sensitivity of the LFA was 99.3%, and the specificity was 98.6%. The results indicated that the specificity, sensitivity, limit of detection, dynamic range, and repeatability of the assay support its use in the field for the purpose of qualitatively evaluating suspicious white powders and environmental samples for the presumptive presence of B. anthracis spores.

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Comparison of Hand-Held Test Kits, Immunofluorescence Microscopy, Enzyme-Linked Immunosorbent Assay, and Flow Cytometric Analysis for Rapid Presumptive Identification of Yersinia pestis

Cited by 31 publications

References 10 publications

Diagnostic Bioluminescent Phage for Detection of Yersinia pestis

Diagnostic Bioluminescent Phage for Detection of Yersinia pestis

Characterization of an F1 Deletion Mutant of Yersinia pestis CO92, Pathogenic Role of F1 Antigen in Bubonic and Pneumonic Plague, and Evaluation of Sensitivity and Specificity of F1 Antigen Capture-Based Dipsticks

Comprehensive Laboratory Evaluation of a Highly Specific Lateral Flow Assay for the Presumptive Identification ofBacillus anthracisSpores in Suspicious White Powders and Environmental Samples

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