Detecting new weapon: curbing drug resistance transmission

Detecting new weapon: curbing drug resistance transmission

November 08, 2016 Source: Science Network

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Researcher Xu Jian, director of the Single Cell Center of the Qingdao Institute of Bioenergy and Process, Chinese Academy of Sciences, told the Journal of the Chinese Academy of Sciences: "Our team has sharply captured the development of drug resistance detection technology in the process of developing new technologies and equipment for microbiology. Potential and opportunity."

Drug resistance quick check problem

At the beginning of the 20th century, one in three people in the world died of pneumonia, tuberculosis, enteritis and diarrhea. According to relevant literature, today's heart disease and cancer have become the main killers of human beings, and the number of people killed by pneumonia and flu is less than 4.5%. This is an important achievement in the field of public health for human application of antibiotics.

Nowadays, human beings have come to the other extreme. The abuse of antibiotics has led to the emergence and widespread spread of drug-resistant bacteria. Anti-drug resistance requires not only the development of new antibiotics, but also the development of drug resistance rapid detection techniques and monitoring systems to improve the targeting and effectiveness of existing antibiotic use, thereby delaying and curbing the spread of drug resistance.

Therefore, on August 26 this year, the National Health and Family Planning Commission and other 14 departments jointly issued the "National Action Plan for Combating Bacterial Resistance (2016-2020)" clearly stated: "Strengthen the application of antibacterial drugs and drug resistance control system" and "improve antibacterial" Drug application and bacterial resistance monitoring system."

According to Xu Jian, since the discovery of bacteria, the culture method is still the mainstream general standard for the susceptibility test of pathogenic bacteria, but for clinical common pathogens, the culture method takes up to 24 to 48 hours (for bacteria that are difficult to grow or grow slowly) There is no way to do it, and at the same time it cannot reveal the mechanism of resistance.

"PCR (polymerase chain reaction) and genome sequencing technology can indirectly speculate the possibility of drug resistance by identifying drug resistance genes." Xu Jian further pointed out that "but the technology relies on the known resistance gene reference sequence, so Unknown resistance cannot be analyzed and is often not quantifiable, so it can only be used as an aid in general."

In order to guide "precise medication" in clinical practice, direct and rapid measurement techniques for bacterial resistance and drug resistance mechanisms are urgently needed. The contradiction between clinical needs and technical status is so urgent and so prominent that on September 8, 2016, the National Institutes of Health offered a reward of 20 million US dollars to specifically stimulate the development of bacterial resistance rapid detection technology.

Raising the concept of "Raman group"

Chinese scientists have been actively trying to explore this field. Since 2012, Xu Jian team has realized the great potential of drug resistance detection technology development. Using the single-cell imaging technology developed by the Single Cell Center in the past six years, Xu Jian and his doctoral student Teng Lin proposed the concept of “Raman group”.

Teng Lin explained to reporters what the “Raman group” is: it is a collection of single-cell Raman spectra of a bacterial cell population under specific conditions and time points. Each single-cell Raman spectrum consists of thousands of Raman peaks corresponding to a class of chemical bonds, reflecting the multidimensional information of the composition and content of specific intracellular chemicals, and its measurement does not require destruction of cells, no labeling, Usually only takes milliseconds or even seconds.

Thus, for any bacterial population, a “Raman group” is equivalent to a “metabolic state” or “metabolite group” that can be measured and monitored at a single cell with precision, and can be directly reflected and characterized for specific Sensitivity and tolerance to drug or environmental changes. Different bacteriostatic mechanisms cause different changes in the intracellular metabolite group, so the "Raman group" changes have the potential to distinguish and even identify various drug stress mechanisms.

Subsequently, a team of more than 10 people from the Single Cell Analysis Department, Bioinformatics Team, and Microbiology Department of the Qingdao Institute of Bioenergy and Process Research of the Chinese Academy of Sciences was established, aiming at the “Raman Group Breaking Through Microbial Resistance Rapid Inspection” .

Everything was difficult at the beginning. During the progress of the project, Teng Lin and her teammates encountered many difficulties, one of which was the data analysis of the “Raman Group”. Teng Lin said: "This series of experiments carried out single-cell Raman imaging on more than 300 cell populations from various drug types, doses, and administration times, and collected Raman spectra of more than 6000 cells, including nearly 9 million. Raman Peak."

Under pressure, this optimistic and determined girl did not bow down and admit defeat. After more than four years of hard work, Teng Lin and other team members of the team developed a series of new methods of data analysis and visualization to dig deeper and verify the interrelationship between the “Raman group” and the microbial drug stress response. This proves that the "Raman group" can quickly measure microbial resistance and distinguish its drug stress mechanism.

Broad application prospects

In addition, the “Raman group” can accurately measure the heterogeneity of drug susceptibility in the bacterial population during drug administration, thus providing a powerful tool for studying the origin and evolution of bacterial resistance. Researchers have demonstrated that the "Raman group" can quickly distinguish between resistant and non-resistant bacteria, so it has the potential to become a new platform technology in both bacteriostatic drug screening and drug-resistant bacterial screening.

Xu Jian said that as a new "phenotype group" means and group big data types, "Raman group" has important advantages. Unlike the traditional transcriptome, proteome or metabolome, the “Raman group” performs rapid and low-cost metabolic state imaging and monitoring of cell populations or communities in a single cell precision, which is a study of the heterogeneity mechanism of life systems. It is of great significance and will have broad application prospects in the fields of drug screening, environmental testing, biological resource mining, and biological process control.

In the detection of drug resistance, the "Raman group" is based on single-cell imaging, independent of bacterial reproduction, and usually can complete bacterial resistance measurement and mechanism differentiation within one hour, so it is in the clinical drug resistance rapid test. Has an important advantage.

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