Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of Phages
The highly pathogenic bacterium <i>Yersinia pestis</i> is the causative agent of plague, a notorious infectious zoonotic disease. When transmitted from person to person as pneumonic plague via droplets, <i>Y. pestis</i> is highly contagious and in most cases is fatal if left...
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MDPI AG
2020-07-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/9/8/611 |
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| author | Friederike Born Peter Braun Holger C. Scholz Gregor Grass |
| author_facet | Friederike Born Peter Braun Holger C. Scholz Gregor Grass |
| author_sort | Friederike Born |
| collection | DOAJ |
| description | The highly pathogenic bacterium <i>Yersinia pestis</i> is the causative agent of plague, a notorious infectious zoonotic disease. When transmitted from person to person as pneumonic plague via droplets, <i>Y. pestis</i> 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. <i>Yersinia pestis</i> specific phages such as L-413C and ΦA1122 are already used for detection of <i>Y. pestis</i> 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 <i>Y. pestis</i>. Genes of putative RBP of phages L-413C (<i>gpH</i>) and ΦA1122 (<i>gp17</i>) were fused with those of fluorescent proteins and recombinant receptor-reporter fusion proteins were produced heterologously in <i>Escherichia coli</i>. When first tested on attenuated <i>Y. pestis</i> 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 <i>Yersinia</i> species and several inactivated fully virulent <i>Y. pestis</i> strains exhibited high specificities of the RBP-reporters against <i>Y. pestis</i>. The L-413C RBP proved to be especially specific, as it only detected <i>Y. pestis</i> at all temperatures tested, whereas the RBP of ΦA1122 also bound to <i>Y. pseudotuberculosis</i> strains at 37 °C (but not at 28, 20 or 6 °C). Finally, the <i>Y. pestis</i>-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. |
| first_indexed | 2024-03-10T18:11:00Z |
| format | Article |
| id | doaj.art-4ad83f7220fa429bbd14974d80fd969f |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T18:11:00Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
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| spelling | doaj.art-4ad83f7220fa429bbd14974d80fd969f2023-11-20T08:05:50ZengMDPI AGPathogens2076-08172020-07-019861110.3390/pathogens9080611Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of PhagesFriederike Born0Peter Braun1Holger C. Scholz2Gregor Grass3Deptartment of Bacteriology and Toxinology, Bundeswehr Institute of Microbiology (IMB), 80937 Munich, GermanyDeptartment of Bacteriology and Toxinology, Bundeswehr Institute of Microbiology (IMB), 80937 Munich, GermanyDeptartment of Bacteriology and Toxinology, Bundeswehr Institute of Microbiology (IMB), 80937 Munich, GermanyDeptartment of Bacteriology and Toxinology, Bundeswehr Institute of Microbiology (IMB), 80937 Munich, GermanyThe highly pathogenic bacterium <i>Yersinia pestis</i> is the causative agent of plague, a notorious infectious zoonotic disease. When transmitted from person to person as pneumonic plague via droplets, <i>Y. pestis</i> 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. <i>Yersinia pestis</i> specific phages such as L-413C and ΦA1122 are already used for detection of <i>Y. pestis</i> 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 <i>Y. pestis</i>. Genes of putative RBP of phages L-413C (<i>gpH</i>) and ΦA1122 (<i>gp17</i>) were fused with those of fluorescent proteins and recombinant receptor-reporter fusion proteins were produced heterologously in <i>Escherichia coli</i>. When first tested on attenuated <i>Y. pestis</i> 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 <i>Yersinia</i> species and several inactivated fully virulent <i>Y. pestis</i> strains exhibited high specificities of the RBP-reporters against <i>Y. pestis</i>. The L-413C RBP proved to be especially specific, as it only detected <i>Y. pestis</i> at all temperatures tested, whereas the RBP of ΦA1122 also bound to <i>Y. pseudotuberculosis</i> strains at 37 °C (but not at 28, 20 or 6 °C). Finally, the <i>Y. pestis</i>-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.https://www.mdpi.com/2076-0817/9/8/611<i>Yersinia pestis</i>plaguemicroscopic assaypathogen detectionreceptor binding proteinphage |
| spellingShingle | Friederike Born Peter Braun Holger C. Scholz Gregor Grass Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of Phages Pathogens <i>Yersinia pestis</i> plague microscopic assay pathogen detection receptor binding protein phage |
| title | Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of Phages |
| title_full | Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of Phages |
| title_fullStr | Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of Phages |
| title_full_unstemmed | Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of Phages |
| title_short | Specific Detection of <i>Yersinia pestis</i> Based on Receptor Binding Proteins of Phages |
| title_sort | specific detection of i yersinia pestis i based on receptor binding proteins of phages |
| topic | <i>Yersinia pestis</i> plague microscopic assay pathogen detection receptor binding protein phage |
| url | https://www.mdpi.com/2076-0817/9/8/611 |
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