Molecular Detection of Helicobacter Pylori: Techniques, Procedures, and Relevance of Bacterial RNA

Helicobacter pylori is a bacterium that infects approximately 4.4 billion people worldwide. Although most infected individuals remain asymptomatic, those who develop symptoms can experience conditions ranging from mild discomfort to severe complications, such as stomach ulcers or the development of certain adenocarcinomas or lymphomas.

Molecular Techniques for Detecting Helicobacter pylori

Various molecular techniques are available for detecting Helicobacter pylori infection, with next-generation sequencing (NGS) and microarrays being the most commonly used. Both techniques require a stomach biopsy as the first step, followed by bacterial RNA extraction and purification. The steps involved are as follows:

1. Sample collection via endoscopy and freezing at -80°C until processing
2. Thawing and tissue lysis:

• In a 2mL tube, add the lysis reagents according to the chosen method, shake for 20 seconds, and centrifuge for 3 minutes at maximum speed in a microcentrifuge.
• Collect the supernatant and store at -80°C

     3. Thawing and RNA purification::

• This involves several sequential steps using various reagents (ethanol, DNase, water, etc.) to isolate the RNA of interest. Between each step, the 2mL tube is shaken and centrifuged for 30 seconds to 2 minutes at 8,000xG.
• The resulting RNA is transferred to a new 2mL tube for subsequent analyses (NGS or microarray)..

Importance of proper RNA purification

Proper purification of bacterial RNA is essential for obtaining reliable and accurate results in diagnosing Helicobacter pylori infection. Chain microcentrifugation plays a crucial role in this process due to its speed and convenience in handling small samples.

To ensure high yields in the total RNA purification of Helicobacter pylori from gastric biopsies, the Biocen 22 equipment stands out as the best option. With a capacity of up to 15,000 RPM and 21,885 xg, it guarantees efficient separation, while its compact design and compatibility with volumes of up to 24 x 2mL ensure precise and reliable sample handling.