Aim: Gene polymorphism biomarkers identify individual susceptibility to environmental and occupational hazards. The conventional approach considers polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis (RFLP), a reliable but expensive and time-consuming two-step procedure. Therefore we evaluated the simpler method confronting two-pair primers (CTPP)-PCR for its robustness and applicability to epidemiologic studies.
Materials & methods: We compared CTPP-PCR and PCR-RFLP techniques to detect two NRF2 polymorphisms in a set of biological samples.
Results: CTPP-PCR produced contradictory results and required the orthogonal technique for confirming the data.
Conclusion: In contrast to PCR-RFLP, CTPP-PCR of NRF2 polymorphisms resulted in ambiguous genotyping which strongly jeopardized heterozygosis classification. The necessity of long optimization and control procedures nullified the potential advantages of CTPP-PCR in terms of costs and time.
Keywords: NRF2; PCR; RFLP; biomarker; confronting two-pair primers; gene polymorphism; oxidative stress; susceptibility.