Table 3.
GJB2 coding sequence mutations detected by SSCP, DHPLC (primer pair Cx26-F-R) and direct sequencing.
| Allele 1 | Allele 2 | Screening techniques | |||||
|---|---|---|---|---|---|---|---|
| Nucleotide change | Amino Acid change | Nucleotide change | Amino Acid change | SSCP | DHPLC | SEQ | |
| 1 | c.551G>C | p.Arg184Pro | - | - | +* | +# | + |
| 2 | c.35delG | Frameshift | c.35delG | Frameshift | + | + | + |
| 3 | c.35delG | Frameshift | c.339T>G | p.Ser113Arg | +/− | + | + |
| 4 | c.101T>C | p.Met34Thr | c.101T>C | p.Met34Thr | + | + | + |
| 5 | c.35delG | Frameshift | c.167delT | Frameshift | +/+ | + | + |
| 6 | c.35delG | Frameshift | c.298C>T | p.His100Tyr | +/+ | + | + |
| 7 | c.457G>A | p.Val153Ile | - | - | + | + | + |
| 8 | c.35delG | Frameshift | c.269insT | Frameshift | +/+ | + | + |
| 9 | c.35delG | Frameshift | c.598G>T | p.Gly200X | +/− | + | + |
| 10 | c.101T>C | p.Met34Thr | - | - | + | + | + |
| 11 | c.167delT | Frameshift | - | - | + | +/− | + |
| 12 | c.79G>A | p.Val27Ile | - | - | − | + | + |
| 13 | c.109G>A | p.Val37Ile | c.109G>A | p.Val37Ile | + | + | + |
| 14 | c.35delG | Frameshift | [c.35delG;c.153delT] | Frameshift | + | + | + |
| 15 | c.35delG | Frameshift | c.94C>T | p.Arg32Cys | +/− | + | + |
| 16 | c.35delG | Frameshift | c.645_648delTAGA | Frameshift | +/+ | + | + |
| 17 | c.269T>C | p.Leu90Pro | c.283G>A | p.Val95Met | +/+ | + | + |
| 18 | c.35delG | Frameshift | c.269insT | Frameshift | +/+ | + | + |
| 19 | c.101T>C | p.Met34Thr | c.298C>T | p.His100Tyr | +/+ | + | + |
| 20 | c.511G>A | p.Ala171Thr | - | - | + | + | + |
| 21 | c.551G>C | p.Arg184Pro | - | - | + | + | + |
| 22 | c.35delG | Frameshift | - | - | + | + | + |
| 23 | c.139G>A | p.Glu47Lys | c.380G>A | p.Arg127His | +/+ | + | + |
| 24 | c.249C>G | p.Phe83Leu | - | - | + | + | + |
| 25 | c.35delG | Frameshift | c.596C>T | p.Ser199Phe | +/− | + | + |
| 27+ | c.35delG | Frameshift | c.71G>A | p.Trp24X | +/− | + | + |
| 28 | c.109G>A | p.Val37Ile | c.109G>A | p.Val37Ile | + | + | + |
| 29 | c.35delG | Frameshift | c.311_324del | Frameshift | +/+ | + | + |
| 30 | c.269T>C | p.Leu90Pro | - | - | + | +/− | + |
| 31 | c.380G>A | p.Arg127His | - | - | + | + | + |
| 32 | c.35delG | Frameshift | c.184G>C | p.Arg184Pro | +/+ | + | + |
| 33 | c.235delC | Frameshift | c.257G>C | p.Ser86Thr | +/+ | + | + |
| 35+ | c.35delG | Frameshift | c.269T>C | p.Leu90Pro | +/+ | + | + |
| 36 | c.235delC | Frameshift | c.558_603dup | Frameshift | +/+ | + | + |
| 37 | c.95G>A | p.Arg32His | c.313_326del | Frameshift | +/− | + | + |
| 38 | c.167delT | Frameshift | c.167delT | Frameshift | + | +/− | + |
| 39 | c.427C>T | p.Arg143Trp | c.313_326del | Frameshift | +/− | + | + |
| 40 | c.109G>A | p.Val37Ile | c.250G>C | p.Val84Leu | +/− | + | + |
| 41 | c.35delG | Frameshift | c.169C>T | p. Gln57X | +/+ | + | + |
| 42 | c.35delG | Frameshift | c.313_326del | Frameshift | +/+ | + | + |
| 43 | c.380G>A | p.Arg127His | - | - | + | + | + |
| 44 | c.109G>A | p.Val37Ile | c.427C>T | p.Arg143Trp | −/− | + | + |
| 45 | c.608T>C | p.Ile203Thr | - | - | + | + | + |
| 46 | c.506G>A | p.Cys169Tyr | - | - | + | + | + |
| 47 | c.479G>A | p.Gly160Ser | - | - | + | +/− | + |
| 49+ | [c.79G>A;c.341A>G] | [p.Val27Ile;p.Glu114Gly] | [c.79G>A;c.341A>G] | [p.Val27Ile;p.Glu114Gly] | −/− | + | + |
| 50 | c.139G>T | p.Glu47X | - | - | + | + | + |
| 51 | c.35delG | Frameshift | c.380G>A | p.Arg127His | +/+ | + | + |
| 52 | c.564_565delGA | Frameshift | - | - | + | + | + |
| 53 | c.109G>A | p.Val37Ile | c.645_648delTAGA | Frameshift | +/− | + | + |
| 54 | c.35G>T | p.Gly12Val | - | - | + | + | + |
| 55 | c.313_326del | Frameshift | - | - | + | + | + |
| Mutation detection accuracy | 82.3% | 98.1 % | 100% | ||||
Novel mutations in bold; SSCP, single-strand conformational polymorphism analysis; DHPLC, denaturing high performance liquid chromatography; SEQ, sequencing
Runs 26, 34 and 48 were wild-type controls
SSCP: +, mutation detected; −, mutation not detected; +/+, both mutations detected; +/−, first mutation detected, second mutation not detected
DHPLC: +, mutation detected in all four runs (each run completed at 3 temperatures); +/−, mutation detected in at least 1 DHPLC run but not in all runs
DNA numbering based on cDNA sequence GenBank NM_004004.3 with +1 as A of ATG start codon.
GJB2-reference: Locus link ID 2706, cDNA AF479776.1; contig NT_024524.13 (1741611..1743919)