BACKGROUND Helicobacter pylori(H.pylori),a globally prevalent pathogen,is exhibiting increasing rates of antimicrobial resistance.However,clinical implementation of pre-treatment susceptibility testing remains limited...BACKGROUND Helicobacter pylori(H.pylori),a globally prevalent pathogen,is exhibiting increasing rates of antimicrobial resistance.However,clinical implementation of pre-treatment susceptibility testing remains limited due to the organism’s fastidious growth requirements and prolonged culture time.AIM To propose a novel detection method utilizing antibiotic-supplemented media to inhibit susceptible strains,while resistant isolates were identified through urease-mediated hydrolysis of urea,inducing a phenol red color change for visual confirmation.METHODS Colombia agar was supplemented with urea,phenol red,and nickel chloride,and the final pH was adjusted to 7.35.Antibiotic-selective media were prepared by incorporating amoxicillin(0.5μg/mL),clarithromycin(2μg/mL),metronidazole(8μg/mL),or levofloxacin(2μg/mL)into separate batches.Gastric antral biopsies were homogenized and inoculated at 1.0×105 CFU onto the media,and then incubated under microaerobic conditions at 37°C for 28-36 hours.Resistance was determined based on a color change from yellow to pink,and the results were validated via broth microdilution according to Clinical and Laboratory Standards Institute guidelines.RESULTS After 28-36 hours of incubation,the drug-resistant H.pylori isolates induced a light red color change in the medium.Conversely,susceptible strains(H.pylori 26695 and G27)produced no visible color change.Compared with the conventional 11-day protocol,the novel method significantly reduced detection time.Among 201 clinical isolates,182 were successfully evaluated using the new method,resulting in a 90.5%detection rate.This was consistent with the 95.5%agreement rate observed when compared with microdilution-based susceptibility testing.The success rate of the novel approach was significantly higher than that of the comparative method(P<0.01).The accuracy of the new method was comparable to that of the dilution method.CONCLUSION The novel detection method can rapidly detect H.pylori drug resistance within 28-36 hours.With its operational simplicity and high diagnostic performance,it holds strong potential for clinical application in the management of H.pylori antimicrobial resistance.展开更多
基金Supported by the Guangxi Science and Technology Major Projects,No.AA23073012the National Natural Science Foundation of China,No.32360035 and No.32060018。
文摘BACKGROUND Helicobacter pylori(H.pylori),a globally prevalent pathogen,is exhibiting increasing rates of antimicrobial resistance.However,clinical implementation of pre-treatment susceptibility testing remains limited due to the organism’s fastidious growth requirements and prolonged culture time.AIM To propose a novel detection method utilizing antibiotic-supplemented media to inhibit susceptible strains,while resistant isolates were identified through urease-mediated hydrolysis of urea,inducing a phenol red color change for visual confirmation.METHODS Colombia agar was supplemented with urea,phenol red,and nickel chloride,and the final pH was adjusted to 7.35.Antibiotic-selective media were prepared by incorporating amoxicillin(0.5μg/mL),clarithromycin(2μg/mL),metronidazole(8μg/mL),or levofloxacin(2μg/mL)into separate batches.Gastric antral biopsies were homogenized and inoculated at 1.0×105 CFU onto the media,and then incubated under microaerobic conditions at 37°C for 28-36 hours.Resistance was determined based on a color change from yellow to pink,and the results were validated via broth microdilution according to Clinical and Laboratory Standards Institute guidelines.RESULTS After 28-36 hours of incubation,the drug-resistant H.pylori isolates induced a light red color change in the medium.Conversely,susceptible strains(H.pylori 26695 and G27)produced no visible color change.Compared with the conventional 11-day protocol,the novel method significantly reduced detection time.Among 201 clinical isolates,182 were successfully evaluated using the new method,resulting in a 90.5%detection rate.This was consistent with the 95.5%agreement rate observed when compared with microdilution-based susceptibility testing.The success rate of the novel approach was significantly higher than that of the comparative method(P<0.01).The accuracy of the new method was comparable to that of the dilution method.CONCLUSION The novel detection method can rapidly detect H.pylori drug resistance within 28-36 hours.With its operational simplicity and high diagnostic performance,it holds strong potential for clinical application in the management of H.pylori antimicrobial resistance.
