Diagnosis of <i>Helicobacter pylori</i> infection

Skrebinska, Sabīne; Mégraud, Françis; Bessède, Emilie

doi:10.1111/hel.12515

Cited by 36 publications

(27 citation statements)

References 39 publications

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“…11 There are several PCR modifications that can help to increase the sensitivity and specificity of detection, including real-time PCR that provides fast detection and quantitation of target DNA. [12][13][14][15] However, this procedure has several disadvantages: high equipment cost, high technical skill, increased risk of false-negative results due to the human error as a result of improper assay development, and incorrect data analysis. 12,16,17 The simpler alternative is nested PCR that involves two rounds of PCR.…”

Section: Introductionmentioning

confidence: 99%

Diagnostic reliability of nested PCR depends on the primer design and threshold abundance ofHelicobacter pyloriin biopsy, stool, and saliva samples

et al. 2020

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Background The aim of this work was to find a reliable nested PCR for the detection of Helicobacter pylori in biopsy, stool, and saliva specimens. Materials and Methods Novel nested PCR was elaborated and validated on 81 clinical biopsy, stool, and saliva samples from the same individual and compared to available H pylori assays: histology, rapid urease test (RUT), stool antigen test (SAT), 13C‐urea breath test (UBT). Results The efficiency and selectivity of 17 published nested polymerase chain reactions (PCR) available for Helicobacter pylori detection were re‐evaluated. Most of them had serious limitations and mistakes in primer design. Hence, we elaborated a nested PCR for the unambiguous identification of H pylori in biopsy, stool, and saliva, using primers targeted to variable regions of the 16S ribosomal RNA (rRNA) gene. Moreover, we determined the detection limit by adding a known number of cells. This number was as low as 0.5 cells in a PCR vial, but due to the DNA isolation procedures, it required 1‐5 × 103 cells/g or ml of specimen. The sensitivity for nested PCR from stomach biopsies was on the same scale as 13C‐UBT (93.8%), but it was much lower in amplifications from stool (31.3%). Sequencing of all obtained PCR products exclusively confirmed H pylori‐specific DNA sequences. Conclusions Elaborated nested PCR assay can serve as an auxiliary method for controversial samples (patients with bleeding or taking proton‐pump inhibitor) in laboratories with basic equipment. The sensitivity and specificity for the amplification from gastric biopsies was almost like 13C‐UBT. Despite the good sensitivity, the threshold occurrence and the ability to survive in the oral cavity aside from and independent of the stomach is the reason why H pylori DNA cannot be reliably detected in saliva, stool, and some biopsy samples.

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Section: Introductionmentioning

confidence: 99%

Diagnostic reliability of nested PCR depends on the primer design and threshold abundance ofHelicobacter pyloriin biopsy, stool, and saliva samples

et al. 2020

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“…Polymerase chain reaction assay of gastric juice has been applied clinically for accurate detection of H pylori . Several genes, including UreA , glmM , UreC , 16S rRNA , 23S rRNA , HSP60 , and VacA genes, have been targeted for PCR analysis .…”

Section: Discussionmentioning

confidence: 99%

“…Polymerase chain reaction assay of gastric juice has been applied clinically for accurate detection of H pylori. 10 Several genes, including…”

Section: Discussionmentioning

confidence: 99%

“…Gastric juiced‐based PCR has been proven to be a reliable method for H pylori diagnosis . It can also evaluate antibiotics resistance and host genetic polymorphisms as well .…”

Section: Introductionmentioning

confidence: 99%

“…Gastric juiced-based PCR has been proven to be a reliable method for H pylori diagnosis. 10 It can also evaluate antibiotics resistance and host genetic polymorphisms as well. 11 However, some of the diagnostic tools such as rapid urease test, culture, and tissue histology have proven to be less accurate when applied to patients with previous eradication failure.…”

Section: Introductionmentioning

confidence: 99%

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Gastric juice‐based PCR assay: An alternative testing method to aid in the management of previously treated Helicobacter pylori infection

et al. 2019

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Background Culture of Helicobacter pylori with previous eradication failure has been emphasized in clinical guidelines. The current unmet need to manage previously treated H pylori is one tool with diagnostic accuracy and ability for antibiotics susceptibility. Gastric juice PCR can provide diagnosis and antibiotics susceptibility; however, whether treatment failure affects its accuracy remains uninvestigated. Our study aimed to investigate diagnostic accuracy and antibiotics susceptibility of juice PCR in previously treated H pylori and to compare with the current standard of culture. Methods We categorized all 547 patients into treatment‐naïve, post‐1st treatment, post‐2nd treatment, and post‐3rd treatment. Helicobacter pylori infection was confirmed using gold standards. Sensitivity, specificity, positive predictive value, negative predictive value, receiver operating characteristic (ROC) curve and area under ROC curve (AUC) of juice PCR and culture were calculated. Intra‐gastric H pylori density was evaluated. Lastly, the antibiotics susceptibility results of gastric juice and culture were compared. Results Our findings demonstrated AUC was higher in juice PCR than culture in all patients (96.7% vs 91.3%, P < 0.0001). The superiority of juice PCR was statistically significant in previously treated patients (P < 0.0001) but not in treatment‐naïve patients (P = 0.13). Antral H pylori density was less marked in previously treated patients (P = 0.014). The comparisons of PCR‐RFLP and E‐test for Clarithromycin resistance showed reliable AUC = 89.8%. Conclusion Compared with the current standard of culture, the gastric juice PCR contains the strengths of performing the antibiotics susceptibility and overcomes the shortcomings of low accuracy. Consequently, gastric juice PCR suits the unmet need to manage previously treated H pylori.

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