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On-Site Fluorescent Detection of Sepsis-Inducing Bacteria using a Graphene-Oxide CRISPR-Cas12a (GO-CRISPR) System.
Kasputis, Tom; He, Yawen; Ci, Qiaoqiao; Chen, Juhong.
Afiliación
  • Kasputis T; Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.
  • He Y; Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.
  • Ci Q; Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.
  • Chen J; Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.
Anal Chem ; 96(6): 2676-2683, 2024 02 13.
Article en En | MEDLINE | ID: mdl-38290431
ABSTRACT
Sepsis is an extremely dangerous medical condition that emanates from the body's response to a pre-existing infection. Early detection of sepsis-inducing bacterial infections can greatly enhance the treatment process and potentially prevent the onset of sepsis. However, current point-of-care (POC) sensors are often complex and costly or lack the ideal sensitivity for effective bacterial detection. Therefore, it is crucial to develop rapid and sensitive biosensors for the on-site detection of sepsis-inducing bacteria. Herein, we developed a graphene oxide CRISPR-Cas12a (GO-CRISPR) biosensor for the detection of sepsis-inducing bacteria in human serum. In this strategy, single-stranded (ssDNA) FAM probes were quenched with single-layer graphene oxide (GO). Target-activated Cas12a trans-cleavage was utilized for the degradation of the ssDNA probes, detaching the short ssDNA probes from GO and recovering the fluorescent signals. Under optimal conditions, we employed our GO-CRISPR system for the detection of Salmonella Typhimurium (S. Typhimurium) with a detection sensitivity of as low as 3 × 103 CFU/mL in human serum, as well as a good detection specificity toward other competing bacteria. In addition, the GO-CRISPR biosensor exhibited excellent sensitivity to the detection of S. Typhimurium in spiked human serum. The GO-CRISPR system offers superior rapidity for the detection of sepsis-inducing bacteria and has the potential to enhance the early detection of bacterial infections in resource-limited settings, expediting the response for patients at risk of sepsis.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones Bacterianas / Técnicas Biosensibles / Sepsis / Grafito Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Humans Idioma: En Revista: Anal Chem Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones Bacterianas / Técnicas Biosensibles / Sepsis / Grafito Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Humans Idioma: En Revista: Anal Chem Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos