CF Gold 1.0 Probe Array CFTR33 ABI CF 2.0 NanoChip® CFTR
Reverse line blot; colorimetric 25 Line probe assay (LiPA); colorimetric 33 OLA; fluorescent 31 Microarray; fluorescent 25
No Yes Yes Yes
Roche Molecular Systems Innogenetics Applied Biosystems Nanogen
NanoChip Workstation NanoChip
Fig. 5. The NanoChip workstation and CF data output (Nanogen Inc., San Diego, CA).
(Roche Molecular Biochemicals, Indianapolis, IN) and the Probe Array CFTR33 (Innogenetics, Inc., Alpharetta, GA) currently screen for 25 and 33 CF mutations, respectively, while also providing optional intron 8 5/7/9T typing (Fig. 3). These reverse line blots or line probe assays (LiPA) are in widespread use because of early availability, ease of use, and ease of validation in the laboratory. The Innogenetics LiPA is also available in a semiautomated platform that minimizes hands-on time for the hybridization and detection steps. Although both assays are amenable to low and high testing volumes, the major disadvantage of these systems is that they are closed platforms, so that additional mutations cannot be tested for easily.
A multiplex oligonucleotide ligation assay (OLA) (ABI, Foster City, CA) and microarray assay (NanoChip®, Nanogen, San Diego, CA) have also been introduced as ASRs. The Nanogen and ABI platforms both offer semiautomated detection and data analysis. Each requires a capital expenditure/reagent rental for instrumentation. The assay formats for both are flexible and can be customized to accommodate additional mutations. Both are amenable to high throughput and can be used as a consolidated work platform for many other assays. OLA is a single - tube non-isotopic test that utilizes automated capillary electrophoresis instrumentation for allele discrimination (Fig. 4). The NanoChip technology offers the latest in allele-specific oligonulceotide hybridization and detection utilizing electronic microarray and fluorescent detection methods (Fig. 5).
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