ABCMdb

PMID: 16379540

Stanziale P, Savino M, De Bonis P, Granatiero M, Zelante L, Bisceglia L
Indirect CFTR mutation identification by PCR/OLA anomalous electropherograms.
Genet Test. 2005 Winter;9(4):285-91., [PubMed]

Sentences

No. Mutations Sentence Comment

2 ABCC7 p.Asp110His ABCC7 p.Ser589Asn Here we report the description of five cases of anomalous electropherograms obtained after PCR/OLA analysis, that led to the identification, in the homozygous state, of two point mutations (D110H and S589N) not included in the assay test panel, a large gene deletion (CFTRdel14b_17b), and an exonic polymorphism (c.4002A Ͼ G). Login to comment 45 ABCC7 p.Asp110His A: Sequencing analysis of exon 4 in case 1 and wild-type control sample. The arrow indicates the nucleotide change leading to the D110H mutation. Login to comment 46 ABCC7 p.Ser589Asn B: Sequencing analysis of exon 12 in case 2 and wild-type control sample. The arrow indicates the nucleotide change leading to the S589N mutation. Login to comment 50 ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Arg1162* ABCC7 p.Gly85Glu ABCC7 p.Ser589Asn FREQUENCY DISTRIBUTION OF CFTR MUTATIONS IDENTIFIED IN 116 PATIENTS WITH CYSTIC FIBROSIS ORIGINATING FROM CENTRAL-SOUTHERN ITALY Mutations Allele frequency (%) F508del 47.41 G542X 9.48 N1303K 5.60 G85E 5.17 2789ϩ5GϾA 1.29 621ϩ1G-ϾT 1.29 R347P 1.29 R553X 1.29 S589N 1.29 W1282X 1.29 CFTRdele14b-17b 0.86 1717-1G-ϾA 0.43 2183 AA-ϾG 0.43 R1162X 0.43 R334W 0.43 711ϩ5G-ϾA 0.43 3849ϩ1OKbC-ϾT 0.43 Unidentified 21.12 A B C D GTTG-3Ј), 14bF (5Ј-GGGAGGAATAGGTGAAGAT-3Ј) and 14bR (5Ј-AATCCACTATGTTTGTATGTA-3Ј), 17bF (5Ј-AA- TGACATTTGTGATATGAT-3Ј) and 17bR (5Ј-ACTTTAG- CTAAGCATTTAAG-3Ј), respectively. Login to comment 59 ABCC7 p.Arg117His ABCC7 p.Tyr122* Case 1 In a 34-year-old male subject affected by obstructive azoospermia resulting from CBAVD diagnosed by scrotal exploration and impalpable vasa clinically observed, no signal could be obtained at either the wild-type or the mutated site with the allele-specific probes R117H, Y122X and 621 ϩ 1G Ͼ T (Fig. 1). Login to comment 62 ABCC7 p.Asp110His This nucleotide transversion generates the D110H mutation (substitution histidine for aspartic acid), often associated with CABVD (Dork et al., 1997). Login to comment 64 ABCC7 p.Arg117His Because the nucleotide substitution occurs 22 bp upstream to the R117H mutation site, it can be postulated that this change does not allow the annealing of the PCR forward primer. Login to comment 66 ABCC7 p.Ser589Asn ABCC7 p.Ser589Asn Sequencing of exon 12 and flanking intronic regions revealed the presence of the nucleotide transition G to A at position c.1898 in the homozygous state generating the substitution of asparagine for serine at codon 589 (S589N) (Fig. 2B). Login to comment 73 ABCC7 p.Arg1162* The presence of signals of wild-type probes of exon 13 (2183AA Ͼ G) and exon 19 (R1162X, 3659delC) mutation sites excluded a deletion involving these exons. Login to comment 94 ABCC7 p.Trp1282* Case 4-5 In two subjects, one oligospermic male and a child suspected of CF because of a borderline sweat test, no signal could be obtained ateither the wild-type or the mutated site, with the allele-specific probes W1282X and 3905insT localized in exon 20 of CFTR gene (Fig. 1D). Login to comment 96 ABCC7 p.Trp1282* This nucleotide change occurred 24 bp downstream from the W1282X site, and we postulated that it prevented amplification of the fragment by affecting the annealing of the reverse PCR primer. Login to comment 118 ABCC7 p.Asp110His ABCC7 p.Ser589Asn In addition, because the D110H and S589N changes, similar to CFTRdel14b_17b, are rare mutations (CF Genetic Analysis Consortium: www.genet.sickkids.on.ca/CFTR) and the c.4002A Ͼ G allelic frequency is 1.32%, as resulted by the analysis performed in 113 control individuals from central-southern Italy (data not shown), it seems to be reasonable to postulate consanguinity in all the families. Login to comment