Multiplex Accelerated PCR System for One-Step Helicobacter pylori cagA + Genotypes Detection: A Guide for Clinical Testing.

Helicobacter pylori cagA + genotype is a leading risk factor for gastric cancer development making accurate identification and timely eradication of H. pylori critical to deadly gastric cancer prevention. Traditional clinical diagnostic methods, including conventional in vitro culture, histological examination, and (13/14)C-urea breath test methods, could only identify the presence of H. pylori, but these means are not capable of identification of cagA + strains. Herein, we firstly built a multiplex detection system based on novel accelerated PCR that could realize one-step detection of as low as 20 copies of H. pylori 16S rDNA and cagA genes within 30 min. In addition, this novel system performed strong anti-jamming capacity, and exhibited that it could specifically differentiate H. pylori cagA- and cagA + genotypes co-existence with other 4 kinds of gastrointestinal pathogens. Furthermore, this one-step system showed remarkable performance on rapid H. pylori infection diagnosis and cagA + genotypes identification in clinical gastric mucosa samples. Specifically, it outperformed histological examination in terms of accuracy and was superior to conventional PCR and DNA sequencing in terms of efficiency. This rapid, sensitive, and reliable H. pylori detection and identification system would break the limitation of traditional methods and realize H. pylori infection diagnosis and cagA + genotypes identification.