ABCMdb

PMID: 15010427

Strom CM, Clark DD, Hantash FM, Rea L, Anderson B, Maul D, Huang D, Traul D, Chen Tubman C, Garcia R, Hess PP, Wang H, Crossley B, Woodruff E, Chen R, Killeen M, Sun W, Beer J, Avens H, Polisky B, Jenison RD
Direct visualization of cystic fibrosis transmembrane regulator mutations in the clinical laboratory setting.
Clin Chem. 2004 May;50(5):836-45. Epub 2004 Mar 9., [PubMed]

Sentences

No. Mutations Sentence Comment

178 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Arg553* ABCC7 p.Arg347Pro ABCC7 p.Gly542* ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Ile506Val The optimal spotting conditions for each probe are indicated by the boxes around spots in C. wild-type controls and heterozygotes for each ACMG mutation and polymorphism, DNA from 12 compound heterozygotes (⌬F508/1898 ϩ 1GϾA, 711 ϩ 1GϾT/⌬F508, G85E/621 ϩ 1GϾT, 3659delC/⌬F508, 3120 ϩ 1GϾA/ 621 ϩ 1GϾT, R347P/G551D, A455E/⌬F508, R560T/ dF508, R553X/⌬F508, 621 ϩ 1GϾT/⌬F508, 621 ϩ 1GϾT/ 711 ϩ 1GϾT, R117H/⌬F508, and I506V/⌬F508) and DNA from 4 homozygous patients (⌬F508 and 2789 ϩ 5GϾA, 3849 ϩ 10kbCϾT, and G542X) was used in validation experiments. Login to comment 181 ABCC7 p.Ile506Val Expected visual spot patterns, as is shown in Fig. 3C, were observed with the exception of a compound heterozygote (I506V/⌬F508) and a homozygous mutation ⌬F508/⌬F508 in exon 10. Login to comment 184 ABCC7 p.Phe508Cys ABCC7 p.Ile506Val ABCC7 p.Ile507Val For example, wild-type probes for I506V and I507V share common sequences with the ⌬F508 probe, and ⌬I507 and F508C have identical wild-type sequence (see Table 2 in the online Data Supplement); this is not surprising because they detect mutations on three contiguous codons. Login to comment 185 ABCC7 p.Phe508Cys ABCC7 p.Ile506Val ABCC7 p.Ile506Val ABCC7 p.Ile507Val ABCC7 p.Ile507Val As a consequence, when the ⌬F508/⌬F508 homozygous mutant sample was tested the F508C, ⌬I507, I506V, and I507V wild-type probes all lost activity, and in the case of the I506V/⌬F508 compound heterozygote, the I507V wild-type probe lost activity. Login to comment 199 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Arg553* ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Arg347Pro ABCC7 p.Gly542* ABCC7 p.Arg560Thr ABCC7 p.Ile507Val In this series, there were 17 ⌬F508 heterozygous patient samples, 1 ⌬F508 homozygous sample, 2 R117H heterozygous samples, and 1 heterozygous patient sample each for I148T, G542X, R553X, R347P, and 2789 ϩ 5GϾA, for a total of 26 mutant alleles. Additional mutant alleles detected in the control samples included three fixed control samples (⌬F508 homozygous, 5T/WT, 3659delC/⌬F508) on every plate and two heterozygous samples (R560T and 1078delT) and one heterozygous sample each for R334W, A455E, R347P, R117H, ⌬I507, I507V, G551D, and 1717-1GϾA as rotating controls. Login to comment 200 ABCC7 p.Asn1303Lys In a second comparison with the Applera CF OLA, Ver. 3.0, we analyzed 1076 samples, including 3 fixed positive controls (⌬F508/⌬F508, 5T/WT, and N1303K/WT) and 1 rotating control of known genotype. Login to comment 203 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Arg117His ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Arg553* ABCC7 p.Arg334Trp ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Arg1162* ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr In this comparison, there were 19 ⌬F508 heterozygous patient samples, 3 I148T heterozygous samples, 3 R117H heterozygous and 1 R117H homozygous samples, 2 W1282X heterozygous samples, and 1 heterozygous patient sample each for G551D, R553X, R1162X, and 3849 ϩ 10kBCϾT, for a total of 36 mutant alleles. Additional mutant alleles detected for this study included fixed controls ⌬F508 homozygous, 5T/WT, and a N1303K heterozygous sample on all plates, and one heterozygous sample each for R560T, G542X, R553X, W1282X, 2184delA, G85E, I148T, 621 ϩ 1GϾT, R334W, R117H, 1078delT, and 1717-1GϾA as rotating controls. Login to comment