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1.
Anal Chim Acta ; 1077: 232-242, 2019 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-31307714

RESUMO

Sepsis is a condition characterized by a severe stage of blood-infection often leading to tissue damage, organ failure and finally death. Fast diagnosis and identification of the sepsis stage (sepsis, severe sepsis or septic shock) is critical for the patient's evolution and could help in defining the most adequate treatment in order to reduce its mortality. The combined detection of several biomarkers in a timely, specific and simultaneous way could ensure a more accurate diagnosis. We have designed a new optical point-of-care (POC) device based on a phase-sensitive interferometric biosensor with a label-free microarray configuration for potential high-throughput evaluation of specific sepsis biomarkers. The sensor chip, which relies on the use of metallic nanostructures, provides versatility in terms of biofunctionalization, allowing the efficient immobilization of different kind of receptors such as antibodies or oligonucleotides. We have focused on two structurally different types of biomarkers: proteins, including C-reactive protein (CRP) and Interleukin 6 (IL6), and miRNAs, using miRNA-16 as an example. Limits of Detection (LoD) of 18 µg mL-1, 88 µg mL-1 and 1 µM (6 µg mL-1) have been respectively obtained for CRP, IL6 and miRNA-16 in individual assays, with high accuracy and reproducibility. The multiplexing capabilities have also been assessed with the simultaneous analysis of both protein biomarkers.


Assuntos
Proteína C-Reativa/análise , Interleucina-6/análise , MicroRNAs/análise , Biomarcadores/análise , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Ouro/química , Limite de Detecção , Análise em Microsséries/instrumentação , Análise em Microsséries/métodos , Nanoestruturas/química , Dispositivos Ópticos , Testes Imediatos , Reprodutibilidade dos Testes , Sepse/diagnóstico
2.
ACS Sens ; 4(1): 52-60, 2019 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-30525470

RESUMO

Existing clinical methods for bacteria detection lack speed, sensitivity, and, importantly, point-of-care (PoC) applicability. Thus, finding ways to push the sensitivity of clinical PoC biosensing technologies is crucial. Here we report a portable PoC device based on lens-free interferometric microscopy (LIM). The device employs high performance nanoplasmonics and custom bioprinted microarrays and is capable of direct label-free bacteria ( E. coli) quantification. With only one-step sample handling we offer a sample-to-data turnaround time of 40 min. Our technology features detection sensitivity of a single bacterial cell both in buffer and in diluted blood plasma and is intrinsically limited by the number of cells present in the detection volume. When employed in a hospital setting, the device has enabled accurate categorization of sepsis patients (infectious SIRS) from control groups (healthy individuals and noninfectious SIRS patients) without false positives/negatives. User-friendly on-site bacterial clinical diagnosis can thus become a reality.


Assuntos
Técnicas Bacteriológicas/métodos , Sangue/microbiologia , Escherichia coli/isolamento & purificação , Interferometria/métodos , Microscopia/métodos , Testes Imediatos , Adsorção , Anticorpos Antibacterianos/química , Anticorpos Antibacterianos/imunologia , Carga Bacteriana/instrumentação , Carga Bacteriana/métodos , Proteínas de Bactérias/química , Técnicas Bacteriológicas/instrumentação , Bioimpressão , Escherichia coli/imunologia , Ouro/química , Humanos , Imunoensaio/instrumentação , Imunoensaio/métodos , Interferometria/instrumentação , Microscopia/instrumentação , Nanoestruturas/química , Análise Serial de Proteínas/instrumentação , Análise Serial de Proteínas/métodos , Sepse/sangue , Sepse/microbiologia
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