Multiplex PCR amplification from the CFTR gene using DNA prepared from buccal brushes/swabs

摘要: Traditionally, DNA used for PCR-based diagnostic analysis has originated from white cells fractionated whole blood. Although this method yields substantial quantities of DNA, there are some drawbacks to the procedure, including inconvenience drawing blood, risk exposure blood-borne pathogens, liquid sample handling, and somewhat involved extraction procedure. Alternatively, genetic diagnosis been derived finger stick blood samples, hair roots, cheek scrapings, urine samples. Oral saline rinses have also extensively as a means collecting buccal epithelial source. However, still requires handling. Herein, we present our results involving rapid collected on cytology brushes swabs use in PCR reactions, specifically multiplex amplification 5 exons within CFTR gene. The quality isolated cells, manner, sufficient reproducibly support amplification. Cheek cell samples prepared them described here highly stable. success rate is 99%. In blind study comparing 12 mutations responsible cystic fibrosis products amplified with both 464 individuals, was 100% correlation validating extracted testing.