Conventional DNA authenticity analyses (RT-PCR) requires samples to be sent to a laboratory. Point-of-use tests, using isothermal amplification, are well characterised but are not in routine use (mainly due to cost and lower sensitivity). This study (purchase required) compared four such amplification techniques for the identification of chicken DNA: loop-mediated isothermal amplification (LAMP), denaturation bubble-mediated strand exchange amplification (SEA), cross-priming amplification (CPA), and recombinase polymerase amplification (RPA). The researchers focussed on the limit of detection, simplicity, amplification time and cost. The LAMP, CPA, and RPA primers all targeted the chicken mitochondrial cytochrome b gene. The SEA primers were provided by the SEA kit. The authors found that all methods showed good specificity to chicken. 0.1% chicken in mutton could be detected using LAMP and RPA methods. The authors considered that, although RPA costs 10 times more than LAMP, the system and primers of LAMP are far more complex. Therefore, they concluded that RPA is the most suitable method in multiplex detection, and LAMP is much better than the other three methods in single-plex detection.
Photo by Braňo on Unsplash
Comments