Characterization of Publicly Available Lymphoblastoid Cell Lines for Disease-Associated Mutations in 11 Genes

Abstract

Clinical genetic testing laboratories offer tests for >800 conditions (1) and need both positive and negative genetic control materials for proficiency evaluation, quality control, and test development/validation. Few such materials are available(2)(3)(4)(5). Genetic material, generally DNA, can be purified from Epstein–Barr virus (EBV)-transformed lymphocyte cell lines. The resulting preparations closely resemble DNA purified from patient samples and often are appropriate controls for clinical genetic testing applications. The National Institute of General Medical Sciences (NIGMS) Human Genetic Cell Repository(6) and the National Institute on Aging (NIA) Aging Cell Repository(7) house thousands of EBV-transformed cell lines, but mutations have been described in only ∼600 of these. We screened 13 cell lines from the NIGMS Repository for medically important mutations in 11 different genes: CFTR , F5 , F2 , MTHFR , HFE , GJB2 (connexin 26), FMR1 (fragile X), HBA1/HBA2 (α-thalassemia), FGFR3 , HD , and HbS/HbC. Six of the cell lines had previously been shown to carry 1 or more mutations in at least one of these genes, and 7 were derived from apparently healthy individuals with no known mutations. Some cell lines were subsequently developed for use as clinically validated positive controls for molecular genetic testing or were used as negative control materials [see accompanying article by Bernacki et al (8), in this issue]. DNA was extracted according to the Puregene protocol (Gentra Systems, Inc.). Selected mutations were analyzed by standard molecular diagnostic techniques at the Duke Molecular Diagnostics Laboratory and/or at other CLIA-certified clinical testing facilities where indicated. The 11 diseases characterized, the specific mutations analyzed, and the general methods used are listed below. The OMIM numbers (9) are listed in Table 1⇓ . #### Cystic Fibrosis (CFTR; ABCC7). Four tests were used to analyze the CFTR gene: the Roche Linear Array CF-31 (Roche …