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

## TL;DR

This study introduces a new high-throughput method to detect Helicobacter pylori's resistance to six antibiotics using multiplex PCR–mass spectrometry mini-sequencing technology.

## Contribution

A novel high-throughput method for detecting H. pylori antibiotic resistance using mPCR-MS mini-sequencing technology.

## Key findings

- The method detected resistance with 99.43% accuracy across 528 results.
- Consistency rates with culture-based testing ranged from 68.2% to 97.0% for different antibiotics.
- The method is extensible and can be improved by adding new mutation sites.

## Abstract

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.