ABCMdb

PMID: 1357180

Cheadle J, Myring J, al-Jader L, Meredith L
Mutation analysis of 184 cystic fibrosis families in Wales.
J Med Genet. 1992 Sep;29(9):642-6., [PubMed]

Sentences

No. Mutations Sentence Comment

13 ABCC7 p.Gly551Asp ABCC7 p.Gly542* '`3 Marked variations in the frequency of AF508 have been reported from different populations.4 For example, in different regions of Europe, Figure 1 The ARMS multiplex analysed on a 3% NuSieve gel. Samples I to 3 are normal, 4 is a AF508 heterozygote, 5 a G542X/AF508, 6 a G551D/AF508, 7 a 621 + IG > T/AF508, and 8 a GSSID homozygote. Login to comment 25 ABCC7 p.Gly551Asp ABCC7 p.Gly542* A multiplex test based on the Amplification Refractory Mutation System (ARMS), developed by Cellmark Diagnostics,'6 allows the simultaneous detection of AF508, G551D, G542X, and 621 + 1G > T. Login to comment 27 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp ABCC7 p.Gly542* ABCC7 p.Gly542* The A reaction identifies normal 621 + 1G> T/ AF508 sequences and mutant G551D/G542X sequences, whereas the B reaction identifies mutant 621 + G> T/AF508 sequences and normal G551D/G542X sequences (fig 1). Login to comment 28 ABCC7 p.Gly551Asp ABCC7 p.Arg553* In parallel to this system we ran a modified version of the multiplex used by Watson et al,'7 which subsequently allowed 621 + IG> T to be screened with AF508, AI507, G551D, and R553X at the same time. Login to comment 31 ABCC7 p.Gly551Asp ABCC7 p.Arg553* A ----239 bp ___186 bp -Heterodup:exes 2 3 4 HincII digestion and analysis on a 10% polyacrylamide gel detects AF508, AI507, G551D, and R553X (fig 2), and an MseI digest followed by analysis on a 3% NuSieve gel detects 621+1IG>T (fig 3). Login to comment 41 ABCC7 p.Gly542* ABCC7 p.Arg1283Met We have identified a total of 89-8% of CF mutations in the Welsh population, with AF508 accounting for 718%, 621+1G>T 6 7%, G542X 2-7%, and R1283M 2-0%. Login to comment 58 ABCC7 p.Gly551Asp ABCC7 p.Arg553* Sample I is normal, 2 a AF508 heterozygote, 3 a AI507 heterozygote, 4 a G551D or R553X heterozygote, and 5 a 621 + IG > T heterozygote. Login to comment 61 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Arg347Pro ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Gln493* ABCC7 p.Val520Phe ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Arg1283Met Welsh Mixed Undefined Total Mutation No % No % No % No % AF508 107/149 71-8 92/126 73 0 69/94 73 4 268/369 72-6 621 + 1G>T 10/42* 6-7 5/34* 4-0 4/25* 4-3 19/101* 51 G551D 2/42* 1-3 6/34* 4-8 3/25* 3-2 11/101* 3 0 G542X 4/42* 2-7 4/34* 3-2 1/25* 1.1 9/101* 2-4 G85E 0/41* 0-0 2/34* 1 6 3/24* 3*4 5/99* 1-4 R553X 2/42* 1-3 2/34* 16 0/25* 00 4/101* 1-1 R1283M 3/42* 2.0 0/34* 0.0 0/25* 0.0 3/101* 0-8 N1303K 1/42* 0 7 1/34* 0-8 0/24* 0.0 2/100* 0-6 AI507 2/149 1-3 0/126 0.0 0/94 0.0 2/369 0-5 R117H 1/42* 0 7 1/34* 0-8 0/25* 0.0 2/101* 0-5 1717- 1G>A 2/42* 1-3 0/34* 0 0 0/25* 0 0 2/101* 0-5 R560T 0/42* 00 0/34* 00 1/25* 1 1 1/101* 03 1154InsTC 0/40* 0 0 1/33* 0 9 0/24* 0.0 1/97* 0-3 V520F 0/42* 0 0 0/34* 0 0 0/25* 0.0 0/101* 0 0 W1282X 0/42* 0 0 0/34* 0.0 0/25* 0.0 0/101* 0 0 R347P 0/42* 0 0 0/34* 0 0 0/24* 0.0 0/100* 0 0 Q493X 0/42* 0 0 0/34* 0 0 0/24* 0 0 0/100* 00 Total (%) 89-8 90 7 86-5 891 * Non-AF508 chromosomes. Login to comment 64 ABCC7 p.Arg1283Met For example 621 + 1G > T is found exclusively on the 1,2,2,2 haplotype, G55 ID is exclusively on the 1,2,2,1 haplotype, and R1283M is always on the 2,2,2,1 haplotype. Login to comment 65 ABCC7 p.Arg553* R553X, however, is associated with two haplotypes (2,1,1,1 and 1,1,2,2) which is consistent with the hypothesis that this is a recurrent mutation.20 Using this correlation, we have attempted to determine the number of mutations that remain unidentified in our total population. Login to comment 75 ABCC7 p.Gly551Asp ABCC7 p.Arg553* A variety of methods for the analysis of multiple cystic fibrosis mutations has been presented.23 After recognising a relatively high frequency of 621 + 1G > T in the Welsh population, we modified a multiplex used by Watson et al'7 to allow the simultaneous detection of this mutation with AF508, AI507, G551D, and R553X; 82-3% of our total CF chromosomes can thus be screened using this system. Login to comment 76 ABCC7 p.Gly551Asp ABCC7 p.Gly542* In parallel to this we ran the ARMS multiplex which detects AF508, G551D, G542X, and 621 + 1G > T, thus accounting for 83-1 % of our total CF chromosomes. Login to comment 80 ABCC7 p.Gly85Glu ABCC7 p.Gly85Glu G85E has been categorised as a severe allele by Zielenski et al.6 However, more recently, Chalkley and Harris24 have reported that G85E may be associated with milder disease (that is, a mild allele). Login to comment 81 ABCC7 p.Gly85Glu In either case, the fact that one of our five AF508/G85E affected subjects is pancreatic sufficient and the other four are insufficient is in conflict with the hypothesis proposed by Kerem et al.3 Table 3 Distribution of XV2c and KM1 9 haplotypes in CF and normal chromosomes. Login to comment 97 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Arg1283Met We wish to thank Professor P S Harper, Dr D Shaw, and Dr A Clarke for their useful comments on this manuscript, Dr L Sandkuijl for his help with the statistical analyses, Dr M 0 621 + 1G> T Total G551D Total G542X Total G85E Total R553X Total R1283M Total N1303K Total AI507 Total RI 17H Total 1717-1 Total R560T Total 1154Ins TC Total 0 1 1 o o o 0 o 0 0 2 0 o o 0 1 0 0 I 1 1 1 I I I Al-Jader, Meredith Table S Distribution of haplotypes in the uncharacterised CF chromosomes. Login to comment 131 ABCC7 p.Arg1283Met A new missense mutation (R1283M) in exon 20 of the cystic fibrosis transmembrane conductance regulator gene. Login to comment 155 ABCC7 p.Gly85Glu A cystic fibrosis patient who is homozygous for the G85E mutation has very mild disease. Login to comment