Application of 16S rRNA gene sequencing in Helicobacter pylori detection

Author:

Szymczak Aleksander1,Ferenc Stanisław2,Majewska Joanna1,Miernikiewicz Paulina1,Gnus Jan3,Witkiewicz Wojciech2,Dąbrowska Krystyna1

Affiliation:

1. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland

2. Regional Specialist Hospital in Wrocław, Research and Development Center, Wrocław, Poland

3. Medical Academy in Wroclaw, Wrocław, Poland

Abstract

Helicobacter pylori is one of the major stomach microbiome components, promoting development of inflammation and gastric cancer in humans. H. pylori has a unique ability to transform into a coccoidal form which is difficult to detect by many diagnostic methods, such as urease activity detection, and even histopathological examination. Here we present a comparison of three methods for H. pylori identification: histological assessment (with eosin, hematoxylin, and Giemsa staining), polymerase chain reaction (PCR) detection of urease (ureA specific primers), and detection by 16S rRNA gene sequencing. The study employed biopsies from the antral part of the stomach (N = 40). All samples were assessed histologically which revealed H. pylori in eight patients. Bacterial DNA isolated from the bioptates was used as a template for PCR reaction and 16S rRNA gene sequencing that revealed H. pylori in 13 and in 20 patients, respectively. Thus, 16S rRNA gene sequencing was the most sensitive method for detection of H. pylori in stomach biopsy samples.

Funder

National Science Centre in Poland

Publisher

PeerJ

Subject

General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

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