Microarray-Based Detection and Clinical Evaluation for Helicobacter pylori Resistance to Clarithromycin or Levofloxacin and the Genotype of CYP2C19 in 1083 Patients

الملخص EN

Background.

Helicobacter pylori (H.

pylori) is one of the most frequent and persistent bacterial infections that affect nearly half of the world’s population.

Antibiotic resistance is a constantly evolving process and local surveillance of antibiotic resistance is warranted to guide clinicians in their choice of therapy.

The aim of this study was to establish a microarray-based detection to identify H.

pylori infection, clarithromycin and levofloxacin susceptibility, and CYP2C19 genetic polymorphism and guide to potential choice of proton pump inhibitor (PPI), antibiotic administration for tailored H.

pylori eradication therapy.

Methods.

By analyzing the sequence of human genomic CYP2C19⁎2 and CYP2C19⁎3 and mutations within the 23S rRNA and gyrA gene regions conferring clarithromycin and levofloxacin resistance, respectively, we developed a microarray for individual therapy detection of H.

pylori infection.

Plasmids were established as positive or limit of detection (LOD) reference materials.

The specificity and sensitivity of the microarray had been performed.

And a total of 1083 gastric biopsy samples were tested and the Kappa value had been calculated between the array and Sanger sequencing.

We also analyzed the resistance to clarithromycin and levofloxacin in China, as well as the CYP2C19 polymorphisms.

Results.

The LOD of detecting H.

pylori was 103 CFU/mL and human genome DNA was 2 ng/μL.

The detection results of 1083 gastric biopsy samples showed that 691 (63.80%) were H.

pylori positive, of which 266 (38.49%) were resistant to clarithromycin, 192 (27.79%) were resistant to levofloxacin, and 61 (8.83%) were resistant to both of them.

For the type of CYP2C19 polymorphism, 412 (38.04%) were homozygous fast type (HomEM), 574 (53%) were heterozygous EM (HetEM), and 97 (8.96%) were poor metabolizer (PM).

Conclusions.

The proposed microarray-based detection has high specificity, sensitivity, and reproducibility for detecting the resistance of clarithromycin or levofloxacin as well as CYP2C19 polymorphism, which may help to improve the clinical eradication rate of H.

pylori.