Abstract
BackgroundHelicobacter pylori (H. pylori) infection is a serious human health threat. The empiric H. pylori treatment paradigm guided by traditional testing technologies has led to antibiotic resistance. Here, we improved the qPCR method to provide technical support for precision H. pylori diagnosis and treatment.MethodsTwo pairs of primers and probes targeting the glmM gene were designed to detect H. pylori, and a multiplex qPCR method was established for virulence factor detection. Then, a rapid urease test (RUT), culturing and qPCR were performed on 141 specimens collected from Xinqiao Hospital of China in 2017 to evaluate the qPCR detection capability. Finally, the H. pylori infectious amount and virulence genes were detected by qPCR.Results1. The improved qPCR method which used two pairs of primers had a higher detection rate (100%) and better accuracy (p = 0.000), compared with the qPCR using a pair of primers. It also had better consistency with the bacterial culture than with RUT (Kappa =0.440, p < 0.001). 2. The H. pylori infectious amount was significantly positively associated with gastritis in corpus (p = 0.003) and gastric erosion (p = 0.043). The H. pylori infectious amount in gastric precancerous patients was significantly lower than that in H. pylori-positive patients (p < 0.05), and the infectious H. pylori-vacA s1+ amount was significantly greater than that of H. pylori-vacA s1- (p < 0.05). 3. The vacA s1 frequency was significantly higher than that of vacA m1/cagA+/babA2+ in chronic superficial gastritis (p = 0.000), peptic ulcer (p = 0.037) and gastric erosion (p = 0.009). The H. pylori-vacA+/cagA+/babA2+ frequency showed a significant positive correlation (p < 0.05).ConclusionsThe H. pylori infectious amount and presence of H. pylori virulence factors showed complex correlations with gastric disease occurrence and development. The improved qPCR with good detection performance can be used for quantitative H. pylori detection and testing for the virulence genes vacA s1, vacA m1, cagA and babA2 simultaneously. These findings will provide valuable information for disease diagnosis and treatment.
Highlights
Helicobacter pylori (H. pylori) infection is a serious human health threat
The improved quantitative real-time polymerase chain reaction (qPCR) with good detection performance can be used for quantitative H. pylori detection and testing for the virulence genes vacA s1, vacA m1, cagA and babA2 simultaneously
Our results suggested that vacA s1 played more important roles in inducing chronic superficial gastritis, peptic ulcer and erosive gastritis than vacA m1, cagA and babA2, which might be partly due to the amount of H. pylori significantly increasing when patients were infected with vacA s1+ strains (p = 0.025)
Summary
Helicobacter pylori (H. pylori) infection is a serious human health threat. The empiric H. pylori treatment paradigm guided by traditional testing technologies has led to antibiotic resistance. Helicobacter pylori (H. pylori) is a bacterial pathogen that infects more than half of the world’s population [1]. The infection is associated with gastrointestinal symptomatology, from gastritis to gastric cancer. Empiric drug use under the guidance of traditional detection technology has caused many problems, such as antibiotic resistance, which is becoming increasingly serious [6,7,8]. In 2019, Barry Marshall suggested that the empiric H. pylori treatment paradigm should be changed to test-guided precision treatment [9]. The transformation of the treatment model is based on the development of precision testing technology
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