Combined rapid susceptibility assay and microorganism identification system

Abstract

In response to the need for highly-sensitive antibiotic susceptibility assays and identification assays that do not require extensive incubation times, the present invention provides automated assay methods and systems that permit the determination of antibiotic susceptibilities and / or microorganism identification in a timeframe that is substantially shorter than has previously been attainable using a hybrid system that combines turbimetric and fluorescence determinations using a single, clear-plastic assay platform. Related devices, kits, and components thereof are also disclosed.

Description

RELATED APPLICATIONS [0001] This application is a continuation of U.S. patent application Ser. No. 09 / 553,223, filed Apr. 20, 2000, the contents of which are hereby incorporated by reference in its entirety, which claims priority of provisional application Ser. No. 60 / 131,829 filed Apr. 29, 1999, entitled “Antibiotic Susceptibility Testing Methods and Devices” and provisional application Ser. No. 60 / 137,819 filed on May 27, 1999, entitled“Antibiotic Susceptibility and Microbial Identification Testing Methods and Systems”.FIELD OF THE INVENTION [0002] A combined anti-microbial susceptibility testing and microorganism identification platform is disclosed. Specifically, the combined anti-microbial susceptibility testing and microorganism identification platform disclosed herein is a hands-free walk-away system ideally suited for clinical microbiology laboratories. More specifically, the automated, combined anti-microbial susceptibility testing and microorganism identification platform ...

AI Technical Summary

Benefits of technology

[0021] It is another object of the present invention to provide a combined rapid anti-microbial susceptibility and microorganism identification system that is fully automated and hand-free.

[0022] It is still another object of the present invention to provide a fully automated, hands free, combined anti-microbial susceptibility and micro-organism identification system that provides, rapid, accurate, and reproducible results in a standardized format.

[0024] The within-disclosed methods differ from those in the art in at least four important respects. First, a hybrid panel concept is utilized, in which samples are assayed via fluorescent as well as turbidimetric means and methods. Second, a modified clear plastic panel is disclosed for the simultaneous assay of samples via fluorescent identification (ID) and turbidimetric antimicrobial susceptibility testing (AST). Third, multiple wavelength optics (e. g. bichromatic calorimetric analysis) are used and applied to enhance sensitivity and accuracy of the assays and systems disclosed herein. Finally, novel methods of determining incubation time and susceptibility results are utilized.

Problems solved by technology

However, life threatening and debilitating systemic and localized microbial infections remains a major healthcare problem.

Mortality resulting from infectious agents remains particularly high among infants, the elderly, the immunosuppressed, patients in long-term care facilities, and skilled nursing homes, Moreover, the emergence of multi-drug resistance organisms such as vancomycin resistant enterococci (VRE) and methacillin resistant Staphylococcus aureaus (MRSA) have increased the challenges of caring for hospitalized patients.

Hospital acquired infections (nosocomial infections) caused by organisms such as VRE, MRSA and pseudomonas add significantly to patient suffering, increased hospital stays, iatrogenic mortality, and increased healthcare costs.

Drug resistance, specifically antibiotic resistance, often occurs when the antibiotic used to treat an infection is either improperly selected, prescribed in a fashion that does not effectively eradicate the infectious agent, or as a result of poor patient compliance.

Furthermore, when ineffective or unnecessary antibiotics are prescribed any infecting bacteria present continues to multiply unabated often resulting in life threatening complications necessitating expensive, aggressive treatments including otherwise needless hospitalization.

However, stains are largely non-specific and seldom definitive.

However, most clinical samples taken directly from infected patients are contaminated and contain PCR inhibitors.

Furthermore, PCR assays are expensive, highly specialized, and require multiple probes in order to identify an unknown organism.

Consequently, PCR is seldom, if ever used to replace standard microbial culture and identification techniques as the front line test in the clinical microbiology laboratory.

Moreover, the labor and material costs associated with such methods are high, consequently only a limited number of biochemical substrates are used in the initial screen.

If the results obtained after the first 18-24 hours of incubation are inconclusive, additional tests must be conducted which further delay definitive results in reaching the physician.

Both the Kirby-Bauer and broth dilution methods are laborious to set up, require skilled personnel to interpret and require a minimum of 18-24 hours before results are available.

The miniaturization of the biochemical assays and drug-containing tubes from 15 mL to 200 μL resulted in difficulty reading plates because visible growth is often barely detectable.

Consequently, much of the time and cost savings associated with the inoculation and set-up was lost reading and reporting results.

This is especially problematic when fluorogenic substrates are used.

In certain cases the growth rate may lag sufficiently so that when a pre-determined reading interval is reached there will be insufficient growth, and hence fluorescence to be detectable.

This in turn could be reported to the clinician that would initiate antimicrobial therapy based on an erroneous result.

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