Droplet Digital PCR Detects Low-Density Infection in a Significant Proportion of Helicobacter Pylori-Negative Gastric Biopsies of Dyspeptic Patients

Ramírez‐Lázaro, María José; Lario, Sergio; Quilez, María Elisa; Montserrat, A.; Bella, María Rosa; Junquera, Félix; García-Martínez, Lorena; Casalots, Àlex; Parra, Tamara; Calvet, Xavier

doi:10.14309/ctg.0000000000000184

Cited by 14 publications

(18 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A recent Spanish study reported that using droplet digital polymerase chain reaction detected low‐density H . pylori infection in 36% of patients with dyspepsia who were diagnosed as negative by routine diagnostic methods 26 …”

Section: Discussionmentioning

confidence: 99%

“…OTUs and RA from 16S rRNA amplicon sequencing method between H. pylorinegative and H. pylori-positive groups stratified based on routine diagnostic methods with dyspepsia who were diagnosed as negative by routine diagnostic methods. 26 Our study was conducted in Mongolia, a country with a high H. pylori prevalence, which increased the statistical power of calculations. However, sample size and power were limited for some of the subgroup analyses.…”

Section: Ta B L E 2 Comparison Of Meanmentioning

confidence: 99%

See 1 more Smart Citation

Advantage of 16S rRNA amplicon sequencing in Helicobacter pylori diagnosis

et al. 2021

View full text Add to dashboard Cite

Background 16S rRNA amplicon sequencing is an accurate method of detecting microbial infection without culture. It is unclear if sequencing has additional benefits over routine diagnostic methods for Helicobacter pylori testing. Methods We enrolled Mongolian volunteers with dyspepsia. Using routine diagnostic methods, positive H. pylori was defined as positive results on histology/immunohistochemistry, culture, rapid urease test, or serology; negative H. pylori was defined by negative results from all these tests. We performed 16S rRNA sequencing on gastric biopsy specimens and calculated cutoffs for operational taxonomic units (OTUs) and relative abundance (RA) to define positive results using ROC curves. Results We examined 161 individuals with a mean age of 43.6 years, and 64.6% were women. Using routine diagnostic methods, 122 (75.8%) participants were H. pylori positive, the sensitivity and specificity for 16S rRNA sequencing were 94.3% and 82.1% or 93.4% and 82.1% when cutoff values were set to 1113 (OTU number) or 4.4% RA, respectively (both p < .001). When combining the validated values, the concordance rate was high (91.1%); however, 16S rRNA sequencing had additional positive yield in 9 cases (5.6%) compared with routine diagnostic methods, and much greater additional positive yield compared to histopathology/IHC, culture, RUT, serology separately with 12 (7.4%), 37 (23.0%) and 43 (26.7%). Conclusion 16S rRNA amplicon sequencing detects potentially important proportion of H. pylori‐positive cases that test negative with routine diagnostic methods. The quantitative number of H. pylori can help to understand how it can be changing by diseases and RA give opportunity to understand how H. pylori communicate with other microbiota.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Ta B L E 2 Comparison Of Meanmentioning

confidence: 99%

Advantage of 16S rRNA amplicon sequencing in Helicobacter pylori diagnosis

et al. 2021

View full text Add to dashboard Cite

show abstract

“…On the aspect of dPCR, studies simultaneously quantified H. pylori clarithromycin-resistant and -susceptible 23S (A2142G, A2142C, and A2143G) and 16s rRNA gene alleles in gastric biopsy and stool samples using droplet digital PCR (ddPCR) and indicated that ddPCR could detect H. pylori and its clarithromycin resistance-associated genotypes and might aid in immediately testing H. pylori status after eradication [126][127][128]. ddPCR was also useful in detecting low-density "occult" H. pylori infection in a significant proportion (36%) of patients diagnosed as negative by conventional methods [129]. Nested dPCR, as another type of dPCR, was also useful to test clarithromycin resistance performed on stool samples in middle school students [130].…”

Section: Recent Advances Of Pcr: Qpcr and Dpcrmentioning

confidence: 99%

Diagnosis of Helicobacter pylori Infection and Recent Advances

Yang

2021

Diagnostics

View full text Add to dashboard Cite

Background: Helicobacter pylori (H. pylori) infects approximately 50% of the world population. Its infection is associated with gastropathies, extra-gastric digestive diseases, and diseases of other systems. There is a canonical process from acute-on-chronic inflammation, chronic atrophic gastritis (CAG), intestinal metaplasia (IM), dysplasia, and intraepithelial neoplasia, eventually to gastric cancer (GC). H. pylori eradication abolishes the inflammatory response and early treatment prevents the progression to preneoplastic lesions. Methods: the test-and-treat strategy, endoscopy-based strategy, and screen-and-treat strategy are recommended to prevent GC based on risk stratification, prevalence, and patients’ clinical manifestations and conditions. Challenges contain false-negative results, increasing antibiotic resistance, decreasing eradication rate, and poor retesting rate. Present diagnosis methods are mainly based on invasive endoscopy and noninvasive laboratory testing. Results: to improve the accuracy and effectiveness and reduce the missed diagnosis, some advances were achieved including newer imaging techniques (such as image-enhanced endoscopy (IEE), artificial intelligence (AI) technology, and quantitative real-time polymerase chain reaction (qPCR) and digital PCR (dPCR). Conclusion: in the article, we summarized the diagnosis methods of H. pylori infection and recent advances, further finding out the opportunities in challenges.

show abstract

“…27 Real-time PCR, dd-PCR, and NGS were also reported to identify low-grade H. pylori infection in a subgroup of patients diagnosed as negative by conventional tests. 18,19,23 Detecting low bacterial load is difficult to achieve but important for us to ensure successful management while avoiding unnecessary treatments. It is meaningful as low-density infection is harmful in some clinical settings, and it may be linked to antibiotic-resistant or more virulent H. pylori strains with certain vacA-cagA genotypes in gastric biopsies.…”

Section: Molecular Diagnosis Of Low-intensity Infection and Verification Of Eradicationmentioning

confidence: 99%

“…pylori diagnosis 16 17 real‐time PCR, 18 droplet digital PCR (dd‐PCR), 19 fluorescent in situ hybridization (FISH), 20 and next‐generation sequencing (NGS) including 16S rRNA amplicon sequencing, 16 transcriptomics, 21 and metagenomics 22 . In this issue, Gantuta et al 23 discussed the advantages of 16S rRNA sequencing in H .…”

Section: Introductionmentioning

confidence: 99%

Application of molecular techniques inHelicobacter pyloridetection: limitations and improvements

Gong

El‐Omar

2021

Helicobacter

View full text Add to dashboard Cite

Application of molecular techniques in Helicobacter pylori detection: limitations and improvements 1 | INTRODUC TI ONHelicobacter pylori (H. pylori), a microaerophilic gram-negative proteobacterium, is the most common human pathogen colonizing the gastric mucosa of over half the world's population. 1,2 Although 70% of infected people are symptomless, H. pylori is recognized as a strong causative factor in many gastrointestinal (GI) diseases such as peptic ulcer, atrophic gastritis, gastric cancer, and mucosa-associated lymphoid tissue (MALT) lymphoma. 3 The transmission of H. pylori is usually through person-to-person contact or from environmental sources via oral-oral, fecal-oral, and gastro-oral routes. 4 Therefore, its infection rate is linked to familial socioeconomic status, overcrowding, poor hygiene, and environmental contamination in food and water supply. 5 While maintaining a spiral rod-shaped form in a favorable environment, H. pylori may transform into coccoid shape, a viable but non-culturable (VBNC) form with low metabolic activity, to survive under stressful conditions including oxygen deprivation, antibiotic exposure, and epithelial attachment. 6 Treatment of the infection is usually a combination of acid inhibitory agents (eg, Proton Pump Inhibitors PPI) and antibiotics (ie, amoxicillin, clarithromycin, metronidazole, and tetracycline), the choice of which depends on antibiotic sensitivity. 7 Efficient diagnosis is required for successful clinical management to relieve symptoms and to eradicate bacteria. Clinicians have developed numerous H. pylori detection methods, which are divided into two groups based on whether samples are collected by invasive endoscopy. 8 Serology, urea breath test (UBT), and stool antigen test (SAT) are the non-invasive tests carried out on stool, breath, saliva, or serum samples. 9 The invasive tests including endoscopy, histology, culture, and rapid urease test (RUT) are carried out on gastric biopsies. 10 All these conventional tests are based on the presence of different components such as enzymes (UBT, RUT), antigens (SAT), bacteria (culture), and antibodies (serology, histology). 11 Several reviews have discussed their advantages and disadvantages in accuracy, sensitivity, specificity, simplicity, and cost. [12][13][14][15] To improve conventional methods, especially sensitivity, some molecular techniques based on DNA/RNA have been recently used in both invasive and non-invasive H. pylori diagnosis. 16 These new methods include polymerase chain reaction (PCR), 17 real-time PCR, 18 droplet digital PCR (dd-PCR), 19 fluorescent in situ hybridization (FISH), 20 and next-generation sequencing (NGS) including 16S rRNA amplicon sequencing, 16 transcriptomics, 21 and metagenomics. 22 In this issue, Gantuta et al. 23 discussed the advantages of 16S rRNA sequencing in H. pylori diagnosis, which brings to attention the need to review molecular detection tests, especially their shortcomings. This editorial summarizes our current understanding of molecular methods, focusing on ...

show abstract

Droplet Digital PCR Detects Low-Density Infection in a Significant Proportion of Helicobacter Pylori-Negative Gastric Biopsies of Dyspeptic Patients

Cited by 14 publications

References 42 publications

Advantage of 16S rRNA amplicon sequencing in Helicobacter pylori diagnosis

Advantage of 16S rRNA amplicon sequencing in Helicobacter pylori diagnosis

Diagnosis of Helicobacter pylori Infection and Recent Advances

Application of molecular techniques inHelicobacter pyloridetection: limitations and improvements

Contact Info

Product

Resources

About