jazyk / language:
Interactive Conference
of Young Scientists
Back to section

”Nested” PCR test for diagnostics of Helicobacter pylori infection


Go to the discussion

”Nested” PCR test for diagnostics of Helicobacter pylori infection

Barbora Šeligová 1 Pavol Sulo 1

1Prírodovedecká fakulta Univerzity Komenského v Bratislave, Bratislava - Karlova Ves, Slovenská republika
seligova12@uniba.sk

According to the European Helicobacter Study Group the main obstacle in the treatment of H. pylori infection is reliable diagnostics. We elaborated “nested“ PCR test, based on the primers designed to the variable regions of 16S rRNA gene, to identify H. pylori in 63 gastric biopsy, 39 saliva and 20 stool samples. The test is 100 times more sensitive than widely used stool antigen test. The reliability was verified by sequencing of 44 PCR products, providing exclusively H. pylori specific DNA sequence. H. pylori was identified in 30% gastric biopsies, in 30% of stool and in 25,6% of saliva samples. The detection limits for individual method was determined by spiking the negative samples with known number of H. pylori cells. In the case of “nested“ PCR it was ≥5x103/g or ml of the sample. The main drawback in non-invasive diagnostic is associated with the low prevalence of H. pylori in stool and saliva (5x103/ml or 5x103/g) in comparison with the gastric biopsies (1x105/g – 2,5x106/g).

Ďakujem Doc. MUDr. Ľudovítovi Lukáčovi, PhD. za poskytnutie vzoriek biopsií a slín, RNDr. Silvii Vaškovej za suspenziu buniek Helicobacter pylori a RNDr. Veronike Fekete za poskytnutie vzoriek stolice.
Bauer B., Meyer T. F. (2011) Ulcers. 2011, p. 1.
Keilberg D., Ottemann K. M. (2016) Environ. Microbiol. 18, p. 791.
Salama N. R., Hartung M. L., Müller A. (2013) Nat. Rev. Microbiol. 11, p. 385.
It appears that your web browser does not support PDF documents viewing

Discussion

Dobrý deň, 02.05.2017 07:33
mňa by zaujímalo z koľkých pacientov ste mali paralelne vzorky stolice, slín aj biopsie? V koľkých prípadoch sa Vám podarilo detegovať H.pylori vo všetkých paralelných vzorkách? Ďakujem
Re: Dobrý deň, 02.05.2017 08:13
k dispozícii sme mali iba paralelné vzorky biopsie a slín, a to presne z 26 pacientov. H. pylori sme detegovali iba v 4 prípadoch z 26. V tomto prípade hovoríme teda o 15% úspešnosti, čo by v prípa...Show all comments
Dobrý deň, 02.05.2017 07:33
aké najmenšie množstvo viete detegovať v slinách?
Re: Dobrý deň, 02.05.2017 08:20
v slinách vieme detegovať najmenej 10 buniek H. pylori v PCR reakcii
Re: Dobrý deň, 02.05.2017 08:20
v slinách vieme detegovať najmenej 10 buniek H. pylori v PCR reakcii
Partners
General partner

Partners
Special thanks
Media partners
Inzercia zadarmo
Organizer