دورية أكاديمية
Culture-Independent Detection and Identification of Leptospira Serovars.
| العنوان: | Culture-Independent Detection and Identification of Leptospira Serovars. |
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| المؤلفون: | Matthias MA; Department of Medicine, Division of Infectious Diseases, University of California, San Diegogrid.266100.3, California, USA.; Department of Internal Medicine, Section of Infectious Diseases, Yale School of Medicine, New Haven, Connecticut, USA., Lubar AA; Department of Medicine, Division of Infectious Diseases, University of California, San Diegogrid.266100.3, California, USA., Lanka Acharige SS; Leptospirosis Research Laboratory, Department of Community Medicine, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Mihintale, Sri Lankagrid.430357.6.; Department of Internal Medicine, Section of Infectious Diseases, Yale School of Medicine, New Haven, Connecticut, USA., Chaiboonma KL; Department of Medicine, Division of Infectious Diseases, University of California, San Diegogrid.266100.3, California, USA., Pilau NN; Department of Medicine, Division of Infectious Diseases, University of California, San Diegogrid.266100.3, California, USA.; Faculty of Veterinary Medicine, Usmanu Danfodiyo University, Sokoto, Nigeria., Marroquin AS; Department of Medicine, Division of Infectious Diseases, University of California, San Diegogrid.266100.3, California, USA.; Department of Internal Medicine, Section of Infectious Diseases, Yale School of Medicine, New Haven, Connecticut, USA., Jayasundara D; Leptospirosis Research Laboratory, Department of Community Medicine, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Mihintale, Sri Lankagrid.430357.6.; Department of Microbiology, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Mihintale, Sri Lankagrid.430357.6., Agampodi S; Leptospirosis Research Laboratory, Department of Community Medicine, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Mihintale, Sri Lankagrid.430357.6.; Department of Internal Medicine, Section of Infectious Diseases, Yale School of Medicine, New Haven, Connecticut, USA.; Department of Community Medicine, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Mihintale, Sri Lankagrid.430357.6., Vinetz JM; Department of Medicine, Division of Infectious Diseases, University of California, San Diegogrid.266100.3, California, USA.; Department of Internal Medicine, Section of Infectious Diseases, Yale School of Medicine, New Haven, Connecticut, USA. |
| المصدر: | Microbiology spectrum [Microbiol Spectr] 2022 Dec 21; Vol. 10 (6), pp. e0247522. Date of Electronic Publication: 2022 Nov 29. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural |
| اللغة: | English |
| بيانات الدورية: | Publisher: ASM Press Country of Publication: United States NLM ID: 101634614 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2165-0497 (Electronic) Linking ISSN: 21650497 NLM ISO Abbreviation: Microbiol Spectr Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, DC : ASM Press, 2013- |
| مواضيع طبية MeSH: | Leptospira*/genetics , Leptospirosis*/diagnosis , Leptospirosis*/veterinary, Humans ; Animals ; Dogs ; Serogroup ; Prospective Studies ; Serum |
| مستخلص: | Pathogenic Leptospira , the causative agents of leptospirosis, comprise >200 serotypes (called serovars). Most have a restricted reservoir-host range, and some, e.g., serovar Copenhageni, are cosmopolitan and of public health importance owing to their propensity to produce severe, fatal disease in humans. Available serotyping approaches-such as multilocus sequence typing, core genome sequence typing, pulsed-field gel electrophoresis, and the cross-agglutination absorption test-are tedious and expensive, and require isolation of the organisms in culture media-a protracted and incredibly inefficient process-precluding their use in prospective studies or outbreak investigations. The unavailability of culture-independent assays capable of distinguishing Leptospira serotypes remains a crucial gap in the field. Here, we have developed a simple yet specific real-time qPCR assay-targeting a Leptospira -unique gene encoding a putative polysaccharide flippase-that provides intraspecies, serotype-defining (i.e., epidemiologically useful) information, and improves upon the sensitivity of preferred lipL32 -based qPCR-based diagnostic tests. The assay, dubbed RAgI ("rage one"), is r apid and a ffordable, and reliably and specifically detects g roup I pathogenic Leptospira in culture, serum, and urine, with no detectable off-target amplification-even of the genetically related but low virulence group II pathogenic (formerly "intermediate") or nonpathogenic Leptospira . It retained 100% diagnostic specificity when tested against difficult sample types, including field-collected dog urine samples and environmental samples containing varied and complex microbial species-consortia. This assay holds considerable promise in the clinical setting, and for routine epidemiological and environmental surveillance studies. IMPORTANCE Leptospirosis is caused by a diverse group of pathogenic spirochetes comprising over 200 different serotypes. Some are widely reported and of public health importance owing to their propensity to produce severe, fatal disease in humans. Apart from their tedium and expense, current serotyping approaches require isolation of the organisms in culture media-a protracted and incredibly inefficient process-rendering them useless clinically and limiting their utilization in prospective studies or outbreak investigations. The unavailability of culture-independent assays capable of distinguishing Leptospira serotypes remains a crucial gap in the field. The 11108 qPCR-assay overcomes this barrier to progress via direct taxonomic and serotype classification of Leptospira from urine and serum samples, and hence, is the first qPCR-based prognostic test for human leptospirosis. |
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| معلومات مُعتمدة: | UL1 TR001863 United States TR NCATS NIH HHS; R21 AI164106 United States AI NIAID NIH HHS; R01 AI108276 United States AI NIAID NIH HHS; U19 AI115658 United States AI NIAID NIH HHS |
| فهرسة مساهمة: | Keywords: Leptospira; classification; diagnostics; leptospirosis; qPCR; serotyping; serum samples; urine samples |
| تواريخ الأحداث: | Date Created: 20221129 Date Completed: 20221227 Latest Revision: 20240205 |
| رمز التحديث: | 20240205 |
| مُعرف محوري في PubMed: | PMC9769591 |
| DOI: | 10.1128/spectrum.02475-22 |
| PMID: | 36445143 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2165-0497 |
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| DOI: | 10.1128/spectrum.02475-22 |