Identification of Cystic Fibrosis Variants by Polymerase Chain Reaction/Oligonucleotide Ligation Assay

Abstract

The purpose of this work is to define rare variants of cystic fibrosis (CF) that are potential sources of error and can confound molecular genetic testing methods. We performed routine , clinical CF mutation screening using a laboratory-developed test and the oligonucleotide ligation assay reagents from Abbott/Celera. In this report, we describe 11 unique allele drop outs [3849 ؉ 10kb C>T (NM_000492.

A Simple Microfluidic Assay for the Detection of Ligation Product

“…There are many practical applications for the detection of ligated products, in addition to the detection of drug resistance-related point mutation, as demonstrated in this study [6].…”

“…A prominent mechanism used to determine the presence of point mutations is through DNA ligation at the position of the mutation [1,3,5,6]. DNA ligases facilitate the joining of nicks on doublestranded DNA duplexes.…”

Newborn cystic fibrosis screening in southeastern Mexico: Birth prevalence and novel CFTR gene variants

“… CF: cystic fibrosis; IRT: immunoreactive trypsinogen; NBS: newborn screening. Complete sequencing of the CFTR gene 29 was performed in all patients, except patient 5, for whom a 32-mutation panel 25 was used. Variant nomenclature at complementary DNA; cDNA and protein levels is according to the reference sequences NM_000531.5 and NP_000522.3, respectively.…”

“…The remaining patient was analyzed against a panel of 32 pathogenic variants (Test Code 10458; Quest Diagnostics Nichols Institute, San Juan Capistrano, CA). 25 Previously unreported missense CFTR variants identified by sequencing were subjected to in silico analysis with the PolyPhen 26 , SIFT, 27 and MutationTaster 28 programs to predict their damaging effects.…”

Cystic Fibrosis Testing Comes of Age

“…The papers by Schwartz et al 14 and Hantash et al 15 fall under the "fine-tuning" category. As robust as the available multiplex testing platforms are, they are still subject to false results due primarily to interfering complexities of the CFTR sequence itself.…”