ABCMdb

PMID: 7684636

Shuber AP, Skoletsky J, Stern R, Handelin BL
Efficient 12-mutation testing in the CFTR gene: a general model for complex mutation analysis.
Hum Mol Genet. 1993 Feb;2(2):153-8., [PubMed]

Sentences

No. Mutations Sentence Comment

48 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* A diird was hybridized with a pool of ASOs for the five most frequent CFTR mutations after AF508: G551D, G542X, W1282X, N1303K, and R553X. Login to comment 50 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr Percent GC content of allele-speciik oligonucleotides GC Content/ASO Mutation G542X G551D R553X W1282X N1303K DI507 1717-1 G - A R560T S549N R117H 621 + 1 G - T 40 53 53 47 41 30 41 53 53 47 30 HS«3X W12»2X N1J0JK £507 H117M • 21 SS4tN RSOOT Table 2. Login to comment 53 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr 1) Number of ASOs (N = 12) 2) Mutation Frequencies # ofCF Mutation Chromosomes Frequency A508 G551D G542X W1282X N1303K R553X 621 + 1 G - T R117H 1717-1 G - A R560T AI507 S549N unknown Total 548 15 13 13 12. Login to comment 54 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Gly542* ABCC7 p.Ser549Asn ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr ABCC7 p.Arg560Thr 9 9 '3 3 2 1 1 133 764 • Minimal Number of Independent Conclusion (4 hybridizations) Filter # 1 N(A5O8) 2 A5O8 72.0% 2.0% 1.7% 2.0% 1.5% 1.1% 1.1% 0.4% 0.4% 0.3% 0.1% 0.1% 17.0% Hybridizations 3 4 G551D 621 + 1 G - T G542X R117H W1282X 1717-1 G - A N13O3K R560T R553X A507 S549N with the remaining six mutations: 621 + 1 G^T, R117H, 1717-1 G - A , R560T, AI507, and S549N. Login to comment 61 ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Arg560Thr These results were scored as 1010 for the sample 2 3 4 5 6 7 8 9 10 N(A5.6) A ( + )Conlrol> 1 2 3 4 5 6 7 8 9 1 0 1 1 1 2 M C o n l r o l s ' 2 3 4 5 6 7 8 * 1 0 1 1 1 2 OS42X Pool GS51D , R553X W 1 2 6 2 X N1303K < 2 3 4 S 8 7 8 9 10 1 1 1 2 /.507 R1 17H Peel 62 1 + 1 2 S54»N R560T 1 7 1 7 - 1 5: in lane 4, rows D and E; 1001 for the sample in lane 3, rows D and E; and 1101 for the sample in lane 5, rows D and E. Login to comment 68 ABCC7 p.Arg553* For example, the pool-1-positive results observed for the sample in lane 4, rows D and E, of Figure 3 was found to be due to the R553X nonsense mutation (Figure 4). Login to comment 69 ABCC7 p.Arg117His In addition, both samples in lanes 3 and 5, rows D and E, of Figure 3 were shown to carry the R117H mutation. Login to comment 70 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp Samples from patients with clinical indications of CF that were positive for only one of the pooled ASOs were then hybridized with the corresponding normal sequence to establish or exclude homozygosity for the mutation (for example, G551D/G551D versus G551D/X). Login to comment 71 ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys lnd«p*nd«nt Hybridizations E A ( B ^ 0 E A ( C* 0 E + )Conlrols * ) C o n l r o l « GS42X GSS1D R553X I W1282X N1303K 1 « Pool 1 Probss 2 3 4 • • 9 • 5 • • A( + ) B B A( + ) B A( + ) B A C ) A 807 R1 17H • 21 + 1 8S4tN RS«0T 1 # f Pool 2 Probes 2 3 4 i 5 » 6 A( B B B A( B A(- 1717-1 Figure 3. Login to comment 72 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Gly542* ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr Example of results obtained from hybridizing four identical filters with ASOs specific for N (the normal sequence at position 508), AF508 (a 3 bp deletion at position 508), pool 1 (G542X, G551D, R553X, W1282X, and N13O3K), and pool 2 (AI507, R117H, 621 + 1 G - T , S549N, R560T, 1717-1 G-A). Login to comment 73 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Arg553* ABCC7 p.Gly542* ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr Row A contains positive control samples with the following genotypes: 1A (AF508/AF508); 2A (AF508/G542X); 3A (AF5O8/G551D); 4A (N/R553X); 5A (NAV1282X); 6A (AF5O8/N13O3K); 7A (N/AI5O7); 8A (N/R117H); 9A (N/621 + 1 G-T); 10A (AF508/S549N); HA (AF508/R560T); and 12A (N/1717-1 G-A). Login to comment 79 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Pool 1: lane 1 (G542X); lane 2 (G551D); lane 3 (R553X); lane 4 (W1282X); lane 5 (N1303K). Login to comment 80 ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr Pool 2: lane 1 (AI507); lane 2 (Rl 17H); lane 3 (621 +1 G-T); lane 4 (S549N); lane 5 (R560T); lane 6 (1717-1 G - A ) . Login to comment 86 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Gly542* ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr Row A contains positive control samples: lane 2 (G542X); lane 3 (G551D); lane 4 (R553X); lane 5 (W1282X); lane 6 (N13O3K); lane 7 (AI507); lane 8 (R117H); lane 9 (621 +1 G-T)); lane 10 (S549N); lane 11 (R560T); lane 12 (1717-1 G-A)). Login to comment 122 ABCC7 p.Arg553* ABCC7 p.Ser549Asn For example, no cross-hybridization is seen here between ASOs for S549N, G55 ID, and R553X, which have identical target locations and differ in sequence by only one base pair. Login to comment