Application of SNaPshot Multiplex Assays for Simultaneous Multigene Mutation Screening in Patients With Idiopathic Sensorineural Hearing Impairment

被引:15
作者
Wu, Chen-Chi [1 ,2 ,3 ]
Lu, Ying-Chang [1 ]
Chen, Pei-Jer [3 ]
Liu, Alyssa Yan-Zhen [5 ]
Hwu, Wuh-Liang [2 ]
Hsu, Chuan-Jen [1 ,4 ]
机构
[1] Natl Taiwan Univ Hosp, Dept Otolaryngol, Taipei 10002, Taiwan
[2] Natl Taiwan Univ Hosp, Dept Med Genet, Taipei 10002, Taiwan
[3] Natl Taiwan Univ, Grad Inst Clin Med, Taipei 10764, Taiwan
[4] Natl Taiwan Univ, Dept Otolaryngol, Taipei 10764, Taiwan
[5] Natl Taiwan Univ, Coll Publ Hlth, Grad Inst Epidemiol, Div Biostat,Coll Med, Taipei 10764, Taiwan
关键词
SNaPshot technique; GJB2; SLC26A4; mitochondrial 12S rRNA; genetic test; NONSYNDROMIC DEAFNESS; TAIWANESE PATIENTS; GENE; IDENTIFICATION; MICROARRAY; PREVALENCE; FEATURES;
D O I
10.1002/lary.20621
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Objectives/Hypothesis: To develop a cost-effective and robust genetic diagnostic tool for patients with idiopathic nonsyndromic sensorineural hearing impairment. Study Design: Development of a diagnostic tool and validation in a prospective cohort. Methods: Twenty common sequence variants in GJB2, SLC26A4, and the mitochondrial 12S rRNA gene were selected based on our previous epidemiological study. These variants were analyzed using the SNaPshot technique. The efficacies of the SNaPshot multiplex assays were determined by using a prospective cohort composed of 214 unrelated Taiwanese patients with idiopathic sensorineural hearing impairment. The results of the assays were compared to the results obtained by direct sequencing. Results: We developed a diagnostic technique consisting of two consecutive panels of SNaPshot multiplex assays, with each panel screening 10 common sequence variants. Theoretically, this design can detect more than 98% of the known deafness-associated sequence variants in Taiwanese individuals. A total of 126 (58.9%) patients were diagnosed as having at least one sequence variant using the SNaPshot multiplex assays. In total, the SNaPshot assays yielded an accuracy of more than 99%. Conclusions: The strengths of SNaPshot multiplex assays include high accuracy, high sensitivity, high flexibility (the examination panel can be easily expanded for additional mutations), low cost (less than US $10 per patient), and easy implementation for any institute with a DNA sequencer. Although only 20 to 30 mutations can be examined in two to three runs of the SNaPshot assay, this technology may be suitable for first-pass screening of deafness-associated mutations in populations with a relatively homogeneous ethnic background.
引用
收藏
页码:2411 / 2416
页数:6
相关论文
共 22 条
[1]   Application of deafness diagnostic screening panel based on deafness Mutation/Gene database using invader assay [J].
Abe, Satoko ;
Yamaguchi, Toshikazu ;
Usami, Shin-Ichi .
GENETIC TESTING, 2007, 11 (03) :333-340
[2]   SUSCEPTIBILITY MUTATIONS IN THE MITOCHONDRIAL SMALL RIBOSOMAL-RNA GENE IN AMINOGLYCOSIDE INDUCED DEAFNESS [J].
BACINO, C ;
PREZANT, TR ;
BU, XD ;
FOURNIER, P ;
FISCHELGHODSIAN, N .
PHARMACOGENETICS, 1995, 5 (03) :165-172
[3]   A rapid method for detection of five known mutations associated with aminoglycoside-induced deafness [J].
Bardien, Soraya ;
Human, Hannique ;
Harris, Tashneem ;
Hefke, Gwynneth ;
Veikondis, Rene ;
Schaaf, H. Simon ;
van der Merwe, Lize ;
Greinwald, John H. ;
Fagan, Johan ;
de Jong, Greetje .
BMC MEDICAL GENETICS, 2009, 10
[4]   A new method for analysis of mitochondrial DNA point mutations and assess levels of heteroplasmy [J].
Cassandrini, D ;
Calevo, MG ;
Tessa, A ;
Manfredi, G ;
Fattori, F ;
Meschini, MC ;
Carrozzo, R ;
Tonoli, E ;
Pedemonte, M ;
Minetti, C ;
Zara, F ;
Santorelli, FM ;
Bruno, C .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2006, 342 (02) :387-393
[5]   Development of a genotyping microarray for Usher syndrome [J].
Cremers, Frans P. M. ;
Kimberling, William J. ;
Kulm, Maigi ;
de Brouwer, Arjan P. ;
van Wijk, Erwin ;
Brinke, Heleen te ;
Cremers, Cor W. R. J. ;
Hoefsloot, Lies H. ;
Banfi, Sandr ;
Simonelli, Francesca ;
Fleischhauer, Johannes C. ;
Berger, Wolfgang ;
Kelley, Phil M. ;
Haralambous, Elene ;
Bitner-Glindzicz, Maria ;
Webster, Andrew R. ;
Saihan, Zubin ;
De Baere, Elfride ;
Leroy, Bart P. ;
Silvestri, Giuliana ;
Mckay, Gareth J. ;
Koenekoop, Robert K. ;
Millan, Jose M. ;
Rosenberg, Thomas ;
Joensuu, Tarja ;
Sankila, Eeva-Marja ;
Weil, Dominique ;
Weston, Mike D. ;
Wissinger, Bernd ;
Kremer, Hannie .
JOURNAL OF MEDICAL GENETICS, 2007, 44 (02) :153-160
[6]   Simultaneous multigene mutation detection in patients with sensorineural hearing loss through a novel diagnostic microarray: A new approach for newborn screening follow-up [J].
Gardner, Phyllis ;
Oitmaa, Eneli ;
Messner, Anna ;
Hoefsloot, Lies ;
Metspalu, Andres ;
Schrijver, Iris .
PEDIATRICS, 2006, 118 (03) :985-994
[7]   Forty-six genes causing nonsyndromic hearing impairment: Which ones should be analyzed in DNA diagnostics? [J].
Hilgert, Nele ;
Smith, Richard J. H. ;
Van Camp, Guy .
MUTATION RESEARCH-REVIEWS IN MUTATION RESEARCH, 2009, 681 (2-3) :189-196
[8]   Mutation spectrum of the connexin 26 (GJB2) gene in Taiwanese patients with prelingual deafness [J].
Hwa, HL ;
Ko, TM ;
Hsu, CJ ;
Huang, CH ;
Chiang, YL ;
Oong, JL ;
Chen, CC ;
Hsu, CK .
GENETICS IN MEDICINE, 2003, 5 (03) :161-165
[9]   Genetic features, clinical phenotypes, and prevalence of sensorineural hearing loss associated with the 961delT mitochondrial mutation [J].
Kobayashi, K ;
Oguchi, T ;
Asamura, K ;
Miyagawa, M ;
Horai, S ;
Abe, S ;
Usami, S .
AURIS NASUS LARYNX, 2005, 32 (02) :119-124
[10]   GENETIC EPIDEMIOLOGIC STUDIES OF EARLY-ONSET DEAFNESS IN THE UNITED-STATES SCHOOL-AGE POPULATION [J].
MARAZITA, ML ;
PLOUGHMAN, LM ;
RAWLINGS, B ;
REMINGTON, E ;
ARNOS, KS ;
NANCE, WE .
AMERICAN JOURNAL OF MEDICAL GENETICS, 1993, 46 (05) :486-491