Multiplex PCR-Mass Spectrometry Mini-Sequencing Technology Detected Antibiotic Resistance of Helicobacter pylori to Six Antibiotics.

The abuse of antibiotics has led to widespread resistance to Helicobacter pylori (H. pylori) in the population. There is an urgent need to establish a method to detect multiple antibiotic resistance rapidly. This study aimed to construct a novel strategy for the high-throughput detection of H. pylori's resistance to varying antibiotics using multiplex PCR-mass spectrometry mini-sequencing (mPCR-MS mini-sequencing) technology. This study detected the resistance of H. pylori to six antibiotics using eight mutated sites (23S rRNA-2143; pbp1A-1667, 1684, 1240; gyrA-261, 271, 573; and 16S rRNA-928) of four resistance genes (pbp1A, gyrA, 23S rRNA, and 16S rRNA), and 525 were detected in all 528 results (99.43%). Then, the culture-based phenotypic drug susceptibility testing (DST) method was used as a reference for drug resistance detection. We found that the consistency rate between mPCR-MS mini-sequencing with the DST results of amoxicillin (AMX), moxifloxacin (MOX), levofloxacin (LEV), clarithromycin (CLA), azithromycin (AZI), and tetracycline (TET) were 95.5% (63/66), 77.3% (51/66), 68.2% (45/66), 93.9% (62/66), 92.4% (61/66), and 97.0% (64/66), respectively. This method was high-throughput and extensible, easily improving the entire detection system by adding new mutation sites. mPCR-MS mini-sequencing technology provides a new approach to mutation sites related to H. pylori's antibiotic resistance.