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. 2003 Oct;41(10):4589-93.
doi: 10.1128/JCM.41.10.4589-4593.2003.

Evaluation of protocol using gene capture and PCR for detection of Helicobacter pylori DNA in feces

W G MacKay  1 , C L Williams, M McMillan, R N Ndip, A J Shepherd, L T Weaver
Affiliations

Evaluation of protocol using gene capture and PCR for detection of Helicobacter pylori DNA in feces

W G MacKay et al. J Clin Microbiol. 2003 Oct.

Abstract

The route of transmission of Helicobacter pylori, which is usually acquired in childhood and is one of the most common bacterial infections in humans, remains undetermined. Mapping the distribution of H. pylori genotypes within families could help to determine the routes of transmission and risk factors. Here we describe a noninvasive method for obtaining H. pylori DNA isolates from the feces of children. Children presenting with gastrointestinal symptoms at the Royal Hospital for Sick Children were tested for gastric H. pylori colonization by using the 13C-urea breath test (UBT) and were asked to provide fecal samples, which were tested for H. pylori by using the HpSA fecal antigen test. DNA was purified from fecal samples by using a novel method of gene capture with subsequent H. pylori PCR analysis. Fifteen UBT-positive and 15 UBT-negative children participated in the study. The positive and negative predictive values for the assay were 80 and 100%, respectively. Fecal DNA purification followed by H. pylori PCR analysis is an effective tool for harvesting H. pylori DNA isolates from the feces of children. This technique may be developed to allow the diagnosis and noninvasive genotyping of H. pylori in children and their families.

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Figures

FIG. 1.
FIG. 1.
Data set showing results for H. pylori PCR compared to the UBT and the HpSA test. The overall positive and negative predictive values for the HpSA and PCR compared to the UBT were 93 and 93%, respectively, and 80 and 100%, respectively.
FIG. 2.
FIG. 2.
Determination of PCR assay specificity and sensitivity. Specificity was tested by using five nonpyloric Helicobacter strains and one C. jejuni strain. Percent differences between the Helicobacter spp. were determined by comparison of 16S rRNA genes as reported by Solnick and Vandamme (16a). The sensitivity of the assay was determined by purification and amplification of serial dilutions of H. pylori cells seeded into fecal slurries from samples from an H. pylori-negative subject. (A) Sensitivity of the assay; (B) specificity of the PCR alone and the assay (DNA purification followed by PCR).
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