Comprehensive Laboratory Evaluation of a Lateral Flow Assay for the Detection of<i>Yersinia pestis</i>

Prentice, Kristin; DePalma, Lindsay; Ramage, J.; Sarwar, Jawad; Parameswaran, Nishanth; Petersen, Jeannine M.; Yockey, Brook; Young, John W.; Joshi, Mrinmayi; Thirunavvukarasu, Nagarajan; Singh, Ajay K.; Chapman, Carol; Avila, Julie R.; Pillai, Christine A.; Manickam, Gowri; Sharma, Shashi; Morse, Stephen; Venkateswaran, Kodumudi Venkat; Anderson, Kevin; Hodge, David R.; Pillai, Segaran P.

doi:10.1089/hs.2019.0094

Cited by 14 publications

(9 citation statements)

References 43 publications

(60 reference statements)

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“…More ideally, rapid and specific confirmatory detection techniques are needed for unambiguous pathogen identification. For Y. pestis , lateral flow immunochromatographic assays employing F1 capsule antibodies have been in successful use for many years and new derivatives are still being developed [ 10 , 31 , 39 ]. For instance, such invaluable rapid tests have been instrumental for on-site plague diagnostics in Madagascar since 2002 [ 40 ].…”

Section: Discussionmentioning

confidence: 99%

Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

Born¹,

Braun²,

Scholz³

et al. 2020

Pathogens

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The highly pathogenic bacterium Yersinia pestis is the causative agent of plague, a notorious infectious zoonotic disease. When transmitted from person to person as pneumonic plague via droplets, Y. pestis is highly contagious and in most cases is fatal if left untreated. Thus, when plague is suspected, rapid diagnosis is crucial, as a serious course of the infection is only averted by early antibiotic therapy. The bacterium is easy to cultivate, accessible and has a high potential for nefarious use such as bioterrorism. Highly specific, rapid and easy-to-use confirmatory diagnostic methods are required to reliably identify the pathogen independently from PCR-based methods or F1 antigen-based immunological detection. Yersinia pestis specific phages such as L-413C and ΦA1122 are already used for detection of Y. pestis in bacterial plaque or biosensor assays. Here, we made use of the host specificities conferred by phage receptor binding (or tail fiber/spike) proteins (RBP) for developing a specific, fast and simple fluorescence-microscopy-based detection method for Y. pestis. Genes of putative RBP of phages L-413C (gpH) and ΦA1122 (gp17) were fused with those of fluorescent proteins and recombinant receptor-reporter fusion proteins were produced heterologously in Escherichia coli. When first tested on attenuated Y. pestis strain EV76, RBP-reporters bound to the bacterial cell surface. This assay could be completed within a few minutes using live or formaldehyde-inactivated cells. Specificity tests using cultures of closely related Yersinia species and several inactivated fully virulent Y. pestis strains exhibited high specificities of the RBP-reporters against Y. pestis. The L-413C RBP proved to be especially specific, as it only detected Y. pestis at all temperatures tested, whereas the RBP of ΦA1122 also bound to Y. pseudotuberculosis strains at 37 °C (but not at 28, 20 or 6 °C). Finally, the Y. pestis-specific capsule, produced when grown at 37 °C, significantly reduced binding of phage ΦA1122 RBP, whereas the capsule only slightly diminished binding of L-413C RBP.

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Section: Discussionmentioning

confidence: 99%

Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

Born¹,

Braun²,

Scholz³

et al. 2020

Pathogens

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“…Rapid BTA assays have previously been evaluated for the detection of biothreat agents including orthopoxviruses, 36 ricin, 37 abrin, 38 Bacillus anthracis, 32,39 and Yersinia pestis. 40 Limited evaluations have also been conducted with assays for the detection of Yersinia pestis, 41 botulinum neurotoxins, 42,43 and staphylococcal enterotoxins. 44 The Tetracore Tularemia BTA test for F. tularensis is available for screening of suspicious powders and/or materials in the field to support necessary public safety actions.…”

Section: Discussionmentioning

confidence: 99%

Comprehensive Laboratory Evaluation of a Specific Lateral Flow Assay for the Presumptive Identification of Francisella tularensis in Suspicious White Powders and Aerosol Samples

et al. 2020

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We conducted a comprehensive, multi-phase laboratory evaluation of the Tularemia BioThreat Alert Ò (BTA) test, a lateral flow assay (LFA) for the rapid detection of Francisella tularensis. The study, conducted at 2 sites, evaluated the limit of detection (LOD) of this assay using the virulent SchuS4 strain and the avirulent LVS strain of F. tularensis. In 6-phase evaluation (linear dynamic range and reproducibility, inclusivity, near-neighbor, environmental background, white powder, and environmental filter extract), 13 diverse strains of F. tularensis, 8 Francisella near neighbors, 61 environmental background organisms, 26 white powders, and a pooled aerosol extract were tested. In the 937 tests performed, the Tularemia BTA demonstrated an LOD of 10 7 to 10 8 cfu/mL, with a sensitivity of 100.00%, specificity of 98.08%, and accuracy of 98.84%. These performance data are important for accurate interpretation of qualitative results arising from screening suspicious white powders in the field.

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“…Alternatively, a positive diagnosis can be determined by observing a 4-fold difference in antibody titers to F1 capsular antigen [54]. Rapid test using ELISA, LFI, PCR, or real-time PCR have been developed to speed up the detection or diagnosis of plague [55][56][57][58][59][60][61][62].…”

Section: Plaguementioning

confidence: 99%

The State of the Art in Biodefense Related Bacterial Pathogen Detection Using Bacteriophages: How It Started and How It’s Going

Sozhamannan

Hofmann

2020

Viruses

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Accurate pathogen detection and diagnosis is paramount in clinical success of treating patients. There are two general paradigms in pathogen detection: molecular and immuno-based, and phage-based detection is a third emerging paradigm due to its sensitivity and selectivity. Molecular detection methods look for genetic material specific for a given pathogen in a sample usually by polymerase chain reaction (PCR). Immuno-methods look at the pathogen components (antigens) by antibodies raised against that pathogen specific antigens. There are different variations and products based on these two paradigms with advantages and disadvantages. The third paradigm at least for bacterial pathogen detection entails bacteriophages specific for a given bacterium. Sensitivity and specificity are the two key parameters in any pathogen detection system. By their very nature, bacteriophages afford the best sensitivity for bacterial detection. Bacteria and bacteriophages form the predator-prey pair in the evolutionary arms race and has coevolved over time to acquire the exquisite specificity of the pair, in some instances at the strain level. This specificity has been exploited for diagnostic purposes of various pathogens of concern in clinical and other settings. Many recent reviews focus on phage-based detection and sensor technologies. In this review, we focus on a very special group of pathogens that are of concern in biodefense because of their potential misuse in bioterrorism and their extremely virulent nature and as such fall under the Centers for Disease and Prevention (CDC) Category A pathogen list. We describe the currently available phage methods that are based on the usual modalities of detection from culture, to molecular and immuno- and fluorescent methods. We further highlight the gaps and the needs for more modern technologies and sensors drawing from technologies existing for detection and surveillance of other pathogens of clinical relevance.

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Comprehensive Laboratory Evaluation of a Lateral Flow Assay for the Detection ofYersinia pestis

Cited by 14 publications

References 43 publications

Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

Comprehensive Laboratory Evaluation of a Specific Lateral Flow Assay for the Presumptive Identification of Francisella tularensis in Suspicious White Powders and Aerosol Samples

The State of the Art in Biodefense Related Bacterial Pathogen Detection Using Bacteriophages: How It Started and How It’s Going

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