On June 22, relying on the original clinical single cell Raman drug sensitivity rapid detection system (cast-r), Chinese researchers established the H.pylori diagnosis and treatment technology cast-r-hp, which integrates the accuracy of a single bacterial cell and the whole process of "identification drug sensitivity traceability". It has the advantages of rapid pathogen identification, accurate drug sensitivity phenotype detection, supporting the research of drug resistance mechanism based on the whole genome of a single cell and accurate traceability. The achievement team is a joint medical, industry, University and research team composed of Qingdao Institute of bioenergy and process, Chinese Academy of Sciences, Institute of infectious diseases prevention and control, Chinese Center for Disease Control and Prevention (Institute of infectious diseases, Chinese Center for Disease Control and prevention), Qingdao Municipal Hospital, Qingdao xingsai biology, etc. the achievement was recently published in clinical chemistry. Helicobacter pylori infection in China is characterized by high infection rate (about 50%), high disease burden, high drug resistance rate and low eradication rate. The main reason for the low eradication rate is the serious drug resistance of Helicobacter pylori strains. Therefore, it is very important for clinical and related disease prevention and control to establish an efficient drug sensitivity diagnosis, treatment and traceability system. At present, the clinical gold standard for the identification of Helicobacter pylori resistant bacteria is the drug sensitivity test combined with endoscopy and bacterial culture. Due to its harsh growth conditions, it usually takes 7-10 days to isolate and culture Helicobacter pylori in gastric biopsy samples, and it is difficult to investigate the "in situ" pathogen diversity in clinical samples. Phenotypic drug sensitivity test after pure culture also takes a long time and requires high technical requirements. However, genotype based drug sensitivity test can only predict the mutations of some known drug resistance genes, and there are different degrees of false negatives. At present, clinicians mainly rely on empirical medication rather than precise treatment under the guidance of drug sensitivity results. However, with the increasing drug resistance of Helicobacter pylori, the eradication failure rate of empirical treatment is becoming more and more serious. In view of the above bottleneck problems, the scientific research team has established a single bacterial cell precision, "identification drug sensitivity traceability" whole process integrated Helicobacter pylori diagnosis and treatment technology cast-r-hp. The reporter learned that the scientific research team has built a single-cell Raman spectrum (SCRs) reference database of 15 common clinical gastric symbiotic bacteria, and then used the end-to-end deep convolution neural network (DCNN) to integrate the Raman spectrum feature extraction with the classification model training process, effectively improving the accuracy of pathogen identification based on SCRs. Helicobacter pylori cells were identified directly from clinical gastric mucosa samples. The accuracy rate was 98.5 ± 0.27%. The researchers established a rapid detection technique for Helicobacter pylori drug sensitivity phenotype to accurately determine the degree of metabolic inhibition by single cell. Compared with the clinical standard E-test method, the drug sensitivity detection accuracy of 17 clinical isolates of Helicobacter pylori to levofloxacin and clarithromycin was 94% and 100% respectively. In addition, the drug sensitivity of Helicobacter pylori cells to levofloxacin and clarithromycin was directly determined in 7 patients' gastric mucosal tissue samples after 48-72 hours of heavy water feeding and drug incubation without isolation and culture. The accuracy was 100%. For the study of drug resistance mechanism and genome-wide traceability, researchers directly identified Helicobacter pylori cells with specific drug sensitivity phenotype from clinical samples, and then completed high balanced target single cell sorting and nucleic acid amplification through Raman optical tweezers droplet single cell sorting (RAGE) chip. After Raman sorting, the whole genome coverage of a single Helicobacter pylori cell can reach 99.7%, and the drug sensitivity phenotype is verified by genotype. It is reported that in the next step, researchers will further develop cast-r-hp system and expand its application based on the "clinical demonstration network of microbial drug sensitive single cell technology", so as to lay a technical and equipment foundation for the establishment of a single cell precision Helicobacter pylori diagnosis, treatment and disease control system. (Xinhua News Agency)
Edit：Li Jialang Responsible editor：Mu Mu
Source：gmw.cn
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