An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i>
Plague, caused by the human pathogen <i>Yersinia pestis</i>, is a severe and rapidly progressing lethal disease that has caused millions of deaths globally throughout human history and still presents a significant public health concern, mainly in developing countries. Owing to the possib...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/11/2/255 |
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| author | Efi Makdasi Yafit Atiya-Nasagi David Gur Ayelet Zauberman Ofir Schuster Itai Glinert Shlomo Shmaya Elad Milrot Haim Levy Shay Weiss Theodor Chitlaru Emanuelle Mamroud Orly Laskar |
| author_facet | Efi Makdasi Yafit Atiya-Nasagi David Gur Ayelet Zauberman Ofir Schuster Itai Glinert Shlomo Shmaya Elad Milrot Haim Levy Shay Weiss Theodor Chitlaru Emanuelle Mamroud Orly Laskar |
| author_sort | Efi Makdasi |
| collection | DOAJ |
| description | Plague, caused by the human pathogen <i>Yersinia pestis</i>, is a severe and rapidly progressing lethal disease that has caused millions of deaths globally throughout human history and still presents a significant public health concern, mainly in developing countries. Owing to the possibility of its malicious use as a bio-threat agent, <i>Y. pestis</i> is classified as a tier-1 select agent. The prompt administration of an effective antimicrobial therapy, essential for a favorable patient prognosis, requires early pathogen detection, identification and isolation. Although the disease rapidly progresses and the pathogen replicates at high rates within the host, <i>Y. pestis</i> exhibits a slow growth in vitro under routinely employed clinical culturing conditions, complicating the diagnosis and isolation. In the current study, the in vitro bacterial growth in blood cultures was accelerated by the addition of nutritional supplements. We report the ability of calcium (Ca<sup>+2</sup>)- and iron (Fe<sup>+2</sup>)-enriched aerobic blood culture media to expedite the growth of various virulent <i>Y. pestis</i> strains. Using a supplemented blood culture, a shortening of the doubling time from ~110 min to ~45 min could be achieved, resulting in increase of 5 order of magnitude in the bacterial loads within 24 h of incubation, consequently allowing the rapid detection and isolation of the slow growing <i>Y. pestis</i> bacteria. In addition, the aerobic and anaerobic blood culture bottles used in clinical set-up were compared for a <i>Y. pestis</i> culture in the presence of Ca<sup>+2</sup> and Fe<sup>+2</sup>. The comparison established the superiority of the supplemented aerobic cultures for an early detection and achieved a significant increase in the yields of the pathogen. In line with the accelerated bacterial growth rates, the specific diagnostic markers F1 and LcrV (V) antigens could be directly detected significantly earlier. Downstream identification employing MALDI-TOF and immunofluorescence assays were performed directly from the inoculated supplemented blood culture, resulting in an increased sensitivity and without any detectable compromise of the accuracy of the antibiotic susceptibility testing (E-test), critical for subsequent successful therapeutic interventions. |
| first_indexed | 2024-03-09T21:15:56Z |
| format | Article |
| id | doaj.art-700c4f2b479d487494f094e9b2a75a24 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-09T21:15:56Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-700c4f2b479d487494f094e9b2a75a242023-11-23T21:32:48ZengMDPI AGPathogens2076-08172022-02-0111225510.3390/pathogens11020255An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i>Efi Makdasi0Yafit Atiya-Nasagi1David Gur2Ayelet Zauberman3Ofir Schuster4Itai Glinert5Shlomo Shmaya6Elad Milrot7Haim Levy8Shay Weiss9Theodor Chitlaru10Emanuelle Mamroud11Orly Laskar12Israel Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelIsrael Institute for Biological Research, Ness-Ziona 74100, IsraelPlague, caused by the human pathogen <i>Yersinia pestis</i>, is a severe and rapidly progressing lethal disease that has caused millions of deaths globally throughout human history and still presents a significant public health concern, mainly in developing countries. Owing to the possibility of its malicious use as a bio-threat agent, <i>Y. pestis</i> is classified as a tier-1 select agent. The prompt administration of an effective antimicrobial therapy, essential for a favorable patient prognosis, requires early pathogen detection, identification and isolation. Although the disease rapidly progresses and the pathogen replicates at high rates within the host, <i>Y. pestis</i> exhibits a slow growth in vitro under routinely employed clinical culturing conditions, complicating the diagnosis and isolation. In the current study, the in vitro bacterial growth in blood cultures was accelerated by the addition of nutritional supplements. We report the ability of calcium (Ca<sup>+2</sup>)- and iron (Fe<sup>+2</sup>)-enriched aerobic blood culture media to expedite the growth of various virulent <i>Y. pestis</i> strains. Using a supplemented blood culture, a shortening of the doubling time from ~110 min to ~45 min could be achieved, resulting in increase of 5 order of magnitude in the bacterial loads within 24 h of incubation, consequently allowing the rapid detection and isolation of the slow growing <i>Y. pestis</i> bacteria. In addition, the aerobic and anaerobic blood culture bottles used in clinical set-up were compared for a <i>Y. pestis</i> culture in the presence of Ca<sup>+2</sup> and Fe<sup>+2</sup>. The comparison established the superiority of the supplemented aerobic cultures for an early detection and achieved a significant increase in the yields of the pathogen. In line with the accelerated bacterial growth rates, the specific diagnostic markers F1 and LcrV (V) antigens could be directly detected significantly earlier. Downstream identification employing MALDI-TOF and immunofluorescence assays were performed directly from the inoculated supplemented blood culture, resulting in an increased sensitivity and without any detectable compromise of the accuracy of the antibiotic susceptibility testing (E-test), critical for subsequent successful therapeutic interventions.https://www.mdpi.com/2076-0817/11/2/255<i>Y. pestis</i>plagueblood cultureF1 and V antigensbacterial diagnostics |
| spellingShingle | Efi Makdasi Yafit Atiya-Nasagi David Gur Ayelet Zauberman Ofir Schuster Itai Glinert Shlomo Shmaya Elad Milrot Haim Levy Shay Weiss Theodor Chitlaru Emanuelle Mamroud Orly Laskar An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i> Pathogens <i>Y. pestis</i> plague blood culture F1 and V antigens bacterial diagnostics |
| title | An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i> |
| title_full | An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i> |
| title_fullStr | An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i> |
| title_full_unstemmed | An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i> |
| title_short | An Improvement in Diagnostic Blood Culture Conditions Allows for the Rapid Detection and Isolation of the Slow Growing Pathogen <i>Yersinia pestis</i> |
| title_sort | improvement in diagnostic blood culture conditions allows for the rapid detection and isolation of the slow growing pathogen i yersinia pestis i |
| topic | <i>Y. pestis</i> plague blood culture F1 and V antigens bacterial diagnostics |
| url | https://www.mdpi.com/2076-0817/11/2/255 |
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