Feasibility of amplification refractory mutation system in fast detection of clarithromycin resistance of Helicobacter pylori in gastric mucosa / 中华消化杂志

ABSTRACT

Objective To evaluate the feasibility of the combination of amplification refractory mutation system (ARMS) and quantitative real-time polymerase chain reaction (PCR) method in fast detection of clarithromycin resistance of Helicobacter pylori (H.pylori) in gastric mucosa.Methods A total of 150 gastric mucosal specimens with positive H.pylori culture were collected from the H.pylori positive patients who failed in H.pylori eradication from January to August in 2013.The drug resistant gene mutation types of H.pylori in these samples were detected by quantitative real-time PCR based on ARMS.And the accuracy was confirmed by sequencing.The clarithromycin resistance of H.pylori was determined by E-assay.Chi-square test was used for statistical analysis.Results Among 149 gastric mucosal specimens (one specimens without wild type or mutation type had been eliminated),the results of quantitative real-time PCR based on ARMS of two samples were not consistent with the results of sequencing;the consistent rate was 98.7％ (147/149).Among 149 specimens with positive H.pylori culture,104 samples (69.8％) were clarithromycin resistance.In 101 samples the clarithromycin resistance was detected by quantitative real-time PCR based on ARMS;the consistent rate was 97.1％ (101/104).Both E-assay and clarithromycin resistant rate detected by E-assay or quantitative real-time PCR based on ARMS was 69.8％ (104/149) and 67.8％ (101/149),respectively,and the difference was not significant (x2 =0.141,P=0.932).Conclusion The combination of ARMS and quantitative real-time PCR method in fast detection of clarithromycin resistance of H.pylori in gastric mucosa is strongly feasible and highly consistent has high consistent rate with sequencing and E-assay.