Selective PCR: a novel internal amplification control strategy for enhanced sensitivity in Salmonella diagnosis.
OBJECTIVE
The aim of this study was to develop a novel strategy that permits the independent amplification of internal amplification control (IAC) and target sequence using the same set of primers, to improve the sensitivity of diagnostic PCR assays.


RESULTS
The method described here is a Salmonella specific PCR test targeting the quorum sensing gene sdiA. It is based on a large size difference between the IAC and the target and consequently on their different extension time. The results indicate the enhanced sensitivity of this test when compared with the competitive IAC strategy. This is demonstrated through parallel testing of artificially contaminated human faecal samples.


CONCLUSIONS
Utilizing this method, the concentration of the IAC, which often leads to false negative results if the target is present in extremely low concentration owing to competition, does not constitute a critical parameter for the detection limit of a PCR assay.


CONCLUSIONS
To our knowledge, this is the first report of using extension time as a critical parameter for the sensitivity of a PCR test. A different approach for the construction of an IAC, based on inverse PCR, has also been introduced.
Oikonomou
I
I
Department of Dietetics and Nutritional Science, Harokopio University, Athens, Greece.
Halatsi
K
K
Kyriacou
A
A
D001426:Bacterial Proteins / Q000235:genetics; D005243:Feces / Q000382:microbiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012015:Reference Standards; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification*; D012480:Salmonella Infections / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D015534:Trans-Activators / Q000235:genetics
D001426:Bacterial Proteins; C097686:SdiA protein, bacteria; D015534:Trans-Activators