ACUSEP

Integrated whole blood acoustophoresis and homogeneous nucleic acid detection cartridge for rapid sepsis diagnostics

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 Obiettivo del progetto (Objective)

'The objective of the ACUSEP research project is to develop an integrated, dry-reagent based disposable ready-to-use cartridge enabling rapid analysis of sepsis-causing organisms and eventual identification of antibiotic resistance directly from suspected blood samples. Sepsis causes annually up to 135 000 deaths in Europe and the early diagnosis is the key to improved survival. Identification of the antibiotic resistance is further beneficial to fast initiation of proper antimicrobial treatment. The rapid diagnostics of sepsis and identification of the causative pathogens can be achieved by research utilizing the recent technological advances; combining acoustophoretic collection of the pathogens from whole blood and photoluminescent dual-tag probe-pair detection technology with nucleic acid amplification providing excellent sensitivity and speed needed in acute diagnostics. The consortium is assembled to cover all the key research areas of the project. It consists of academic research organizations with significant technical and scientific experience of acoustophoretic target organism collection and purification, nucleic acid amplification, photoluminescent reporters and multiplexed homogeneous probe detection technologies. The project will proceed in phases, divided to both parallel and serial tasks in collaboration with SME and clinical partners involved, to perform a clinical evaluation of the developed prototype consumable assay cartridge and to initiate further exploitation activities. The specific objective in FP7 HEALTH-2010 addressed to is to “develop detection and analytical tools and technologies for diagnosis, monitoring and prognosis of diseases.'

Introduzione (Teaser)

Sepsis causes annually up to 135 000 deaths in Europe, and rapid diagnosis is the key to improved survival. Combining pathogen collection from whole blood using ultrasonic sound waves and photoluminescent DNA detection technology promises to make this a reality.

Descrizione progetto (Article)

Current diagnostic methods for sepsis rely on culturing patient blood and tests to identify bacteria and their antibiotic resistance, which take up to several days. Rapid diagnostics of sepsis-causing bacteria can save the lives of infected patients.

The EU-funded http://www.acusep.fi (ACUSEP) project aimed to provide a proof-of-concept for new sepsis diagnostics that can detect very few bacteria in the blood within hours. The objective of ACUSEP was to develop a disposable ready-to-use cartridge enabling rapid analysis of sepsis-causing organisms directly from suspected blood samples.

To achieve this, scientists used the microfluidic method of acoustophoresis to collect bacteria by ultrasonic sound waves in a small sample. Polymerase chain reaction (PCR) was used to identify bacteria and antibiotic resistance genes.

The detection system consists of several components developed by project participants. Acoustophoretic devices for separation and enrichment of bacteria from blood proved to work with blood samples spiked with bacteria. Novel lanthanide luminescence label molecules detected bacterial DNA in a PCR reaction. Solid-phase oligonucleotide probe arrays allowed for homogeneous multiplexed detection of PCR amplification products. Sensitive PCR assays detected and identified the five most important sepsis-causing bacteria along with several antibiotic resistance genes.

In the integrated system PCR assays, converted to dry-reagent chips allowed automated processing of the PCR reaction. The project fabricated and tested four successive prototypes of the integrated system. The most successful prototype was tested in a clinical study and compared with traditional sepsis diagnostics. The ACUSEP prototype detected bacteria in the blood of patients within two hours. The results were later confirmed by blood culture.

ACUSEP's system has high potential. If developed further and processed to a commercial product, it could become important in the care of suspected sepsis patients in hospitals. Rapid identification of the sepsis causing bacteria and the possible antibiotic resistance they carry will decrease mortality and morbidity.