Normalized real-time PCR for diagnosis of H. pylori infection

Ebtisam E. Al-Moayad, Saeed M. Alghalibi, Hassan A. Al-Shamahy, Akram T. Nasher, Nezar N. Alhebshi

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Objectives: There is increasing interest in the use of quantitative PCR (q-PCR) for diagnosis of H. pylori infection. However, the assay remains largely unstandardized, making comparison between studies unreliable. The objective of this study was to assess accuracy of a normalized q-PCR assay for diagnosis of the infection.

Subjects and methods: Seventy-six fresh gastric biopsy specimens were collected from patients undergoing upper gastrointestinal tract endoscopy and examined by rapid urease test (RUT), culture, and a commercial TaqMan q-PCR assay targeting the ureA gene. Counts obtained from the latter assay were normalized to the human ACTB gene. A subject was considered to be infected if two or more assays were positive.

Results: The detection rates were 42.1%, 52.6%, and 78.9% by culture, RUT and q-PCR, respectively. Bacterial density ranged 0.005 to 4800 bacteria per 100 human cells. Because q-PCR showed low initial specificity (45.7%), the cutoff value for the assay was recalculated as 1 bacterium per 100 human cells, using ROC curve analysis. Accordingly, the sensitivities and specificities were 79.5% and 97.3%, respectively, for culture; 94.9% and 91.9%, respectively, for RUT; and 94.9% and 94.6%, respectively, for q-PCR. By gold standard, 39 of the dyspeptic patients (51.3%) were found to be infected.

Conclusions: With the identified cutoff value, the q-PCR assay diagnosed H. pylori infection with an accuracy slightly superior to that of RUT. However, the possibility that low counts detected only by q-PCR represent true infections warrants further investigation. Normalization of bacterial counts for standardization of q-PCR H. pylori assays is recommended.

Original languageEnglish
Pages (from-to)123-9
Number of pages7
JournalQatar Medical Journal
Volume2014
Issue number2
DOIs
StatePublished - Dec 1 2014

Keywords

  • Culture
  • Helicobacter pylori
  • Normalization
  • Quantitative PCR
  • Rapid urease test

Fingerprint

Dive into the research topics of 'Normalized real-time PCR for diagnosis of H. pylori infection'. Together they form a unique fingerprint.

Cite this