Helicobacter pyloriIs Not Eradicated after Triple Therapy: A Nested PCR Based Study

Abstract

Detection of Helicobacter pylori after triple therapy is usually carried out by either rapid urease test (RUT), urea breath test (UBT), histology, bacterial isolation, and single round PCR or serological tests. In this study, antral biopsy specimens from 25 patients were tested for H. pylori by RUT, culture, histology, and nested PCR in their antral biopsy specimens before and after treatment. Three genes, namely, heat shock protein (hsp60), phosphoglucosamine mutase (ureC), and flagellar export ATP synthase (fliI) of H. pylori were targeted. Of the 25 antral biopsy specimens, the RUT, culture, histology, and nested PCR positivity dropped from 81.8% to 12%, 31% to 0%, 100 to 84%, and 100% to 92%, respectively, before and after therapy. Further, hsp60 specific amplicons from 23 out of 25 patients gave identical restriction pattern, while 6 fliI and 1 ureC specific amplicon produced different restriction pattern. Furthermore, variations in fliI gene sequences in H. pylori after treatment were also confirmed by sequencing and compared in silico. Nested PCR based detection of H. pylori is more sensitive method to detect H. pylori after therapy than culture, RUT, and histology. Further, this study suggests that H. pylori is not eradicated completely after triple therapy.

Possible association between Helicobacter pylori infection and vocal fold leukoplakia

“…In the current study, a highly conserved gene, HSP60, was targeted to identify H. pylori by nested PCR, which has been proposed as the gold standard test . There was a higher positive rate of H. pylori detected by nested PCR compared with the single‐step PCR and rapid urease tests, in line with the previous study . Fang et al and Siupsinskiene et al found that 39.4% and 32% of vocal polyps cases were H. pylori ‐positive using the rapid urease test, respectively.…”

“…13,27 There was a higher positive rate of H. pylori detected by nested PCR compared with the single-step PCR and rapid urease tests, in line with the previous study. 14 Fang et al 28 and Siupsinskiene et al 9 found that 39.4% and 32% of vocal polyps cases were H. pylori-positive using the rapid urease test, respectively. Although, in our study, we discovered a low H. pylori-positive rate of 11.4% in vocal fold polyp.…”

“…Each of them has one or more advantages or disadvantages. Nonetheless, the nested PCR‐based technique, if performed properly, could be proposed as the gold standard test …”

“…Nonetheless, the nested PCR-based technique, if performed properly, could be proposed as the gold standard test. 13,14 The purpose of this study was to investigate whether H. pylori can colonize in the vocal folds of patients with vocal fold leukoplakia. It is the first study to determine the relationship between H. pylori infection and vocal fold leukoplakia.…”

Gastritis – facts and doubts

“…Only the PCR method allows the detection of even very few bacteria present in specimens of mucosa. Thus, in the study by Patel et al , 4 weeks after eradication with a triple combination drug therapy, H. pylori was detected in only 16% of patients on the histopathological examination and in 12% on the urease test, while the PCR method revealed the presence of the same bacterial strains in as many as 92% of patients [31]. In these patients such an insidious infection can support an inflammatory state then wrongly considered to be H. pylori negative.…”