Application of the Luminex LabMAP in Rapid Screening for Mutations in the Cystic Fibrosis Transmembrane Conductance Regulator Gene: A Pilot Study

Abstract

Cystic fibrosis (CF) is an autosomal recessive disorder that occurs in ∼1 in 2500 Caucasians (1)(2)(3). CF can result from combinations of >750 known mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, and in certain populations, as many as 1 person in 25 is a carrier (1)(2)(3). To date, screening methods have either been narrow, requiring multiple methodologies to be brought to bear for comprehensive coverage, or are cost prohibitive (1)(4). This study describes the Luminex LabMAPTM system and its potential for simultaneous, rapid, sensitive, and specific screening for mutations in the CFTR gene. The LabMAP system incorporates polystyrene microspheres that are internally dyed with two spectrally distinct fluorochromes. Using precise ratios of these fluorochromes, an array is created consisting of 100 different microsphere sets with specific spectral addresses. Each microsphere set can possess a different reactant on its surface. Because microsphere sets can be distinguished by their spectral addresses, they can be combined, allowing up to 100 different analytes to be measured simultaneously in a single reaction vessel. A third fluorochrome coupled to a reporter molecule quantifies the biomolecular interaction that has occurred at the microsphere surface. Microspheres are interrogated individually in a rapidly flowing fluid stream as they pass by two separate lasers in the Luminex100 analyzer. High-speed digital …