Kb. Bamford et al., NESTED PRIMERS IMPROVE SENSITIVITY IN THE DETECTION OF HELICOBACTER-PYLORI BY THE POLYMERASE CHAIN-REACTION, The Journal of infection, 36(1), 1998, pp. 105-110
To investigate potential routes of spread of infection by the polymera
se chain reaction (PCR) it is important that the technique is effectiv
e in the types of specimen to be investigated. To establish the limits
of detection of Helicobacter pylori by PCR in clinical material from
the gastric mucosa, faeces, dental plaque and oral rinses, samples wer
e seeded with known numbers of bacteria. DNA extraction was followed b
y amplification with primers from the urease C gene. Nested primers we
re used to amplify the PCR product which was detected using a digoxige
nin-labelled probe. Faeces or plaque inhibited the single reaction 10(
2)-10(6) fold, A second amplification using nested primers and probing
increased the sensitivity to a level similar to that obtained with pu
re culture. This method is potentially useful with less likelihood of
false negative results when trying to detect H. pylori by PCR in highl
y contaminated, clinical material.