Projects / Programmes
Metrology for monitoring nosocomial respiratory tract infections caused by Gram-negative bacteria
| Code |
Science |
Field |
Subfield |
| 2.15.00 |
Engineering sciences and technologies |
Metrology |
|
| Code |
Science |
Field |
| P180 |
Natural sciences and mathematics |
Metrology, physical instrumentation |
| Code |
Science |
Field |
| 2.02 |
Engineering and Technology |
Electrical engineering, Electronic engineering, Information engineering |
Metrology
Metrological support for diagnostics
Gram-negative bacteria
Nosocomial infection
Respiratory tract infection
Antimicrobial resistance
Researchers (1)
| no. |
Code |
Name and surname |
Research area |
Role |
Period |
No. of publicationsNo. of publications |
| 1. |
37410 |
PhD Alexandra Bogožalec Košir |
Biology |
Head |
2019 - 2021 |
0 |
Organisations (1)
| no. |
Code |
Research organisation |
City |
Registration number |
No. of publicationsNo. of publications |
| 1. |
0105 |
National Institute of Biology |
Ljubljana |
5055784 |
0 |
Abstract
Bacterial infections are a major cause of hospitalization, with 5-10% of hospitalizations resulting in nosocomial infection in Europe and North America and more than 40% in South America, Sub‑Saharan Africa and Asia. Five out of six bacteria that most commonly cause nosocomial infection, are Gram-negative bacteria. They are responsible for 87% of hospital-acquired pneumonia, 45–70% of ventilator-associated pneumonia, 20–30% of catheter-related bloodstream infections, and commonly cause other intensive care unit complication.
Bacterial cultures and other standard microbiological techniques, have been predominantly used for detection and identification of bacteria. However, cultivation and phenotypic tests take days and delay introduction of infection control measures. Additionally, as the conventional methods are unable to identify the causative agent quickly enough, infection is consequently often treated empirically with broad-spectrum antibiotics. This can lead to the emergence of antimicrobial resistance. Accurate diagnosis, rapid tracing of outbreak source, as well as robust approaches for assessing key epidemiological indicators, such as disease prevalence and incidence, and monitoring of drug resistance, is thus crucial. The needs for traceability and standardisation of nucleic acid measurements, together with the need for metrological support of measurements, including those for antimicrobial resistance have already been recognised on different levels, from research to state, and the importance of the metrological aspects have been emphasised in international projects (e.g. INFECT-MET, AntiMicroResist, SEPTIMET).
To enable reliable implementation of new technologies, they should be assessed in terms of accuracy and sensitivity. At the moment, higher order methods and reference materials that would provide reference for such assessments are lacking. The main objective of the proposed research is to develop and evaluate higher order methods for accurate measurement of Gram-negative bacteria causing nosocomial respiratory infection, and their susceptibility/resistance to antibiotics, and value assignment of calibrators or reference materials in order to establish a reference measurement system to support faster molecular diagnostics in the presented field.
First a literature survey of nucleic-acid amplification-based methods for detection/quantification of most frequent Gram-negative bacteria causing nosocomial respiratory tract infections, complete with any potential resistance mechanisms will be done. Relevant molecular methods will be selected and in-silico evaluated in terms of specificity and applicability and further in vitro evaluated according to several performance criteria such as sensitivity, specificity, robustness, repeatability, and reproducibility. Measurement uncertainty will be determined. For the development and evaluation of the methods control materials of different levels of complexity will be developed and evaluated in terms of possible use as candidate reference materials. As quality and quantity of nucleic acids is highly dependent on extraction method, a literature survey will be done to find and select most relevant extraction methods, which will be evaluated using whole organisms. Developed methods will be evaluated in terms of possible use as reference measurement procedures and used to assess the influence on clinical diagnostics. Based on performance, potential higher order method will be proposed including extraction and detection/quantification of nucleic acid.
The proposed research will bring fuller understanding of the need for the metrological support in clinical setting and contribute to the prevention of unnecessary use of antibiotics in patients with illnesses, such as acute respiratory-tract infections. The principle of development of reference methods and materials as well as principles of evaluation of emerging approaches will be transferable to other clinically relevant organism
Significance for science
Methods currently being performed for detection and quantification of Gram-negative bacteria, including their susceptibility/resistance to antibiotics will be better supported through the application of metrological considerations and the application of traceable methods that are currently lacking. The results gained with this study will serve as a basis for further reference method and material development and metrological aspect of the methods will serve as support to diagnostic laboratories in evaluation of other detection approaches. Additionally, the collected data on Gram-negative bacteria present in nosocomial respiratory tract infections will provide foundation for broader epidemiological studies.
The proposed project represents a breakthrough in the concept of understanding metrological support needed to underpin nucleic-acid (DNA and RNA) amplification-based methods for detection of Gram-negative bacteria causing nosocomial respiratory tract infection. The results of this project will have important implications for currently used and emerging methods, as candidate reference measurement procedures and candidate reference materials will be developed, which can be used for evaluation of performance of currently used method and the assessment of the potential of new emerging molecular methods. Moreover, the metrology being proposed in this project will be essential for maximising the potential of the new emerging molecular tools for monitoring clinical and environmental samples.
The research proposed in this project will thus be of great importance for further recourses for standardisation of infectious disease monitoring. Additionally, investigation of the metrology requirements for emerging molecular approaches for surveillance purposes will also have an impact on antimicrobial resistance testing, as more specific and sensitive methods enable more precise treatment with the appropriate antibiotics.
Significance for the country
Methods currently being performed for detection and quantification of Gram-negative bacteria, including their susceptibility/resistance to antibiotics will be better supported through the application of metrological considerations and the application of traceable methods that are currently lacking. The results gained with this study will serve as a basis for further reference method and material development and metrological aspect of the methods will serve as support to diagnostic laboratories in evaluation of other detection approaches. Additionally, the collected data on Gram-negative bacteria present in nosocomial respiratory tract infections will provide foundation for broader epidemiological studies.
The proposed project represents a breakthrough in the concept of understanding metrological support needed to underpin nucleic-acid (DNA and RNA) amplification-based methods for detection of Gram-negative bacteria causing nosocomial respiratory tract infection. The results of this project will have important implications for currently used and emerging methods, as candidate reference measurement procedures and candidate reference materials will be developed, which can be used for evaluation of performance of currently used method and the assessment of the potential of new emerging molecular methods. Moreover, the metrology being proposed in this project will be essential for maximising the potential of the new emerging molecular tools for monitoring clinical and environmental samples.
The research proposed in this project will thus be of great importance for further recourses for standardisation of infectious disease monitoring. Additionally, investigation of the metrology requirements for emerging molecular approaches for surveillance purposes will also have an impact on antimicrobial resistance testing, as more specific and sensitive methods enable more precise treatment with the appropriate antibiotics.
