Centre Technology Transfer CITTRU

A new PCR method for detection of bacteria and fungi in biological sample.

Posted by Centre Technology Transfer CITTRUResponsive · Innovative Products and Technologies · Poland

Summary of the technology

In the treatment of bloodstream infections the most important and difficult problem of determining the effectiveness of treatment and, consequently, the cost and time of hospitalization is the effective diagnosis of factors causing a systemic inflammatory response in sepsis. The material subjected to diagnostic testing is blood taken from a patient showing clinical signs of sepsis. The greatest difficulty is the very small amount of microorganisms being responsible for the infection in the blood, or they are only periodically seeded into the blood. Currently, the standard diagnostic blood cultures are performed with specialized broth media, preferably in automatic culture systems. A weakness of these methods are time consumption (until a result of the study) and low sensitivity, which causes that in only 15-20% of the culture it is possible to achieve the growth of microorganisms. The situation is further complicated by the fact that patients undergo antibiotic treatment before a blood sample is taken for culture. In this case blood cultures are very difficult due to the fact that they contain antibiotics which inhibit the growth of microorganisms. The molecular biology techniques, such as PCR or hybridization, belong to diagnostic methods that provide reliable, accurate and fast diagnosis of bloodstream infections. The sensitivity of molecular methods is much higher than the sensitivity of the culture method. However, currently available systems enable the detection of several particular species of microorganisms or every possible gene in theory, but on the other hand they require sequencing of PCR product, which in turn increases the cost and time for the result.
The proposed new PCR method allows simultaneous DNA detection of the entire panel of bacterial and fungal microorganisms, with the differentiation of Gram-negative bacteria, Gram-positive bacteria, yeasts fungi and molds in a sample of biological material (including patient's spittle or blood). There is the possibility of using the detection method only for bacteria or fungi, or for the simultaneous detection of both fungi and bacteria, which reduces the cost of testing. There is also the opportunity to identify a specific species of microorganism after sequencing of the PCR product.

Description of the technology

The offer is a new PCR method for the simultaneous detection of bacteria and fungi in a sample of biological material. This technology allows simultaneous DNA detection of Gram-negative bacteria, Gram-positive bacteria, yeast fungi and molds in a sample of biological material, including patient’s spittle or blood. Thus it finds an application in the biomedical field.

Specifications

The offered invention relates to a new method for detection of bacteria and fungi in a sample of biological material, in which the contained DNA is amplified by real-time multiplex PCR system. An amplification is carried out in two stages. In the first stage there are used primers specific for bacteria and fungi. Then the product of first amplification is used as a template in a second step – an amplification using primers and probes to differentiate yeast fungi and molds, Gram-positive and Gram-negative bacteria.

Main advantages of its use

  • The main advantages of the proposed method are:• detection of the entire panel of bacterial and fungal microorganisms, with the differentiation of Gram-negative bacteria, Gram-positive bacteria, yeasts fungi and molds, but without typing a particular species,• simultaneous amplification of at least two DNA sequences, which allows a simultaneous detection of bacteria and fungi, without waiting for the result of DNA electrophoresis,• application of a nested-PCR system which allows for increased sensitivity of detection by two orders of magnitude compared to one-stage PCR;• the possibility of using the detection method only for bacteria or fungi, or for the simultaneous detection of both fungi and bacteria, which reduces the cost of testing;• the opportunity to identify a specific species of microorganism after sequencing of the PCR product.

Applications

  • The proposed method allows simultaneous DNA detection of Gram-negative bacteria, Gram-positive bacteria, yeast fungi and molds in a sample of biological material, including patient’s spittle or blood. Thus it finds an application in the biomedical field.

Related keywords

  • Microbiology Technology
  • Microbiology Market
  • bloodstream infection
  • sepsis
  • PCR
  • bacteria
  • fungi
  • biological sample

About Centre Technology Transfer CITTRU

Technology Transfer Office from Poland

Centre for Innovation, Technology Transfer and University Development (CITTRU) is a part of Jagiellonian University, whose role is to promote university research, to support innovation and to create cooperation with the business. CITTRU main task is to offer the scientific achievements of the Jagiellonian University in the market by providing legal protection, licensing, sale of intellectual property rights, creation of academic business, coordination of company-ordered research projects, etc. Currently promoted technologies are mainly focused on new materials science, pharmacology and medical technology.

Inventions offered by Jagiellonian University are promoted and awarded during numerous exhibitions, e.g. 58th International Exhibition of Innovation, Research and New Technologies INNOVA (BRUSSELS 2009), 38th International Exhibition of Invention New Techniques & Products (Geneva 2010) or 24th International exhibition of environmental equipment, technologies and services POLLUTEC (Paris 2009).

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