Sensitive and efficient detection of RB1 gene mutations enhances care for families with retinoblastoma

被引:209
作者
Richter, S
Vandezande, K
Chen, N
Zhang, K
Sutherland, J
Anderson, J
Han, LP
Panton, R
Branco, P
Gallie, B
机构
[1] Univ Toronto, Dept Ophthalmol, Toronto, ON M5S 1A1, Canada
[2] Univ Toronto, Dept Mol & Med Genet, Toronto, ON M5S 1A1, Canada
[3] Univ Toronto, Dept Med Biophys, Toronto, ON M5S 1A1, Canada
[4] Univ Toronto, Univ Hlth Network, Vis Sci Res Program, Toronto, ON M5S 1A1, Canada
[5] Univ Toronto, Hosp Sick Children, Toronto, ON M5G 1X8, Canada
[6] Univ Toronto, Retinoblastoma Solut, Toronto, ON, Canada
[7] Univ Western Ontario, Fac Med, Toronto, ON, Canada
关键词
D O I
10.1086/345651
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Timely molecular diagnosis of RB1 mutations enables earlier treatment, lower risk, and better health outcomes for patients with retinoblastoma; empowers families to make informed family-planning decisions; and costs less than conventional surveillance. However, complexity has hindered clinical implementation of molecular diagnosis. The majority of RB1 mutations are unique and distributed throughout the RB1 gene, with no real hot spots. We devised a sensitive and efficient strategy to identify RB1 mutations that combines quantitative multiplex polymerase chain reaction (QM-PCR), double-exon sequencing, and promoter-targeted methylation-sensitive PCR. Optimization of test order by stochastic dynamic programming and the development of allele-specific PCR for four recurrent point mutations decreased the estimated turnaround time to <3 wk and decreased direct costs by one-third. The multistep method reported here detected 89% (199/224) of mutations in bilaterally affected probands and both mutant alleles in 84% (112/134) of tumors from unilaterally affected probands. For 23 of 27 exons and the promoter region, QM-PCR was a highly accurate measure of deletions and insertions (accuracy 95%). By revealing those family members who did not carry the mutation found in the related proband, molecular analysis enabled 97 at-risk children from 20 representative families to avoid 313 surveillance examinations under anesthetic and 852 clinic visits. The average savings in direct costs from clinical examinations avoided by children in these families substantially exceeded the cost of molecular testing. Moreover, health care savings continue to accrue, as children in succeeding generations avoid unnecessary repeated anaesthetics and examinations.
引用
收藏
页码:253 / 269
页数:17
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