A Rapid and High-throughput Multiplex Genetic Detection Assay for Detection, Semi-quantification and Virulence Genotyping of in Non-invasive Oral Samples

Overview

Journal Front Cell Infect Microbiol

Specialties Cell Biology
Infectious Diseases
Microbiology

Date 2023 Oct 16

PMID 37842005

Authors

Wenjing Chi

Su Wang

Tao Liu

Wenrong Jiang

Li Ding

Yingxin Miao

Feng Yang

Jinghao Zhang

Danian Ji

Zili Xiao

Haowei Zhu

Yong Wu

Zhijun Bao

Hu Zhao

Shiwen Wang

Affiliations

Soon will be listed here.

Abstract

Aim: This study established a high-throughput multiplex genetic detection assay (HMGA) for rapid identification, semi-quantification and virulence analysis of directly from the clinical non-invasive oral samples.

Methods: The gastric mucosa and oral samples were collected from 242 patients in Shanghai from 2021 to 2022. All the samples were detected by routine clinical tests for and Sanger sequenced for inconsistent results. A new multiplex PCR assay providing results within 4 hours was designed and optimized involving fluorescent dye-labeled specific primers targeted gene, semi-quantitative gene and 10 virulence genes of . Semi-quantification was carried out by simulating the serial 10-fold dilutions of positive oral samples, and the loads in different clinical samples were further compared. The mixed plasmids of virulence genes , and were used to evaluate the performance on different genotypes. The consistency of 10 virulence genes in gastric mucosa, saliva, mouthwash and dental plaque of -positive patients was compared.

Results: The non-invasive HMGA was highly specific for detection of all 12 targets of and human internal reference gene , and the sensitivity to all target genes could reach 10 copies/μL. Compared with routine clinical tests and sequencing, non-invasive HMGA has a high level (>0.98) of sensitivity, specificity, accuracy, PPV, NPV and kappa coefficient for direct detection of in oral samples. Moreover, by detecting peak area levels of , it was confirmed that the loads in gastric mucosa were significantly higher than those of the three kinds of oral samples (<0.05). We also found that 45.0% (91/202) of patients had different virulence genes in different oral samples. The concordance of positive detection rates of each virulence gene between saliva and gastric mucosa was more than 78% (<0.05).

Conclusion: The non-invasive HMGA proved to be a reliable method for the rapid identification, semi-quantification and detection of 10 virulence genes directly in oral samples, providing a new idea for non-invasive detection of .

Citing Articles

Helicobacter pylori in oral cavity: current knowledge.

Costa L, Carvalho M, Vale F, Marques A, Rasmussen L, Chen T Clin Exp Med. 2024; 24(1):209.

PMID: 39230790 PMC: 11374826. DOI: 10.1007/s10238-024-01474-1.

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