A comparison of restriction fragment length polymorphism, tetra primer amplification refractory mutation system PCR and unlabeled probe melting analysis for LTA+252 C > T SNP genotyping

作者：Soler Stephan; Rittore Cecile; Touitou Isabelle; Philibert Laurent^*

来源：Clinica Chimica Acta, 2011, 412(5-6): 430-434.

DOI：10.1016/j.cca.2010.11.012

• SNP detection
• RFLP
• TPAP
• UPMA

Background: From the wide range of methods currently available for genotyping, we wished to identify a quick, reliable and affordable approach for routine use in our laboratory for LTA + 252 C>T SNP screening.
Methods: We set up and compared three genotyping methods for SNP detection: restriction fragment length polymorphism (RFLP), tetra primer amplification refractory mutation system PCR (TPAP) and unlabeled probe melting analysis (UPMA). The SNP model used was LTA + 252 C>T, a cytokine gene polymorphism that has been associated with response to treatment in rheumatoid arthritis. The study was performed using 46 samples from healthy Caucasian volunteers.
Results: Allele and genotype distribution was similar to that previously described in the same population. All three genotyping methods showed good reproducibility and are suitable for a medium scale throughput molecular platform. UPMA was the most cost effective, reliable and safe method since it required the shortest technician time, could be performed in a single closed tube and involved automatic data analysis.
Conclusion: This work is the first to compare these three genotyping techniques and provides evidence for UPMA being the method of choice for LTA + 252 C>T SNP genotyping.

• 出版日期2011-2-20