$(this).attr("href","#"); // //$(this).bind('click', function() { SetNumTwo(33618)}); // var url = "../userInformation/PDFLogin.aspx"; // var refererrurl = document.referrer; // var downloadurl = window.location.href; // var args = "PaperID=" + 33618 + "&RefererUrl=" + refererrurl + "&DownloadUrl=" + downloadurl; // url = url + "?" + args + "&rand=" + RndNum(4); // // $(this).bind('click', function() { ShowTwo(url)}); // } // }); // } // //获取下载pdf注册的cookie // function getcookie() { // var cookieName = "pdfddcookie"; // var cookieValue = null; //返回cookie的value值 // if (document.cookie != null && document.cookie != '') { // var cookies = document.cookie.split(';'); //将获得的所有cookie切割成数组 // for (var i = 0; i < cookies.length; i++) { // var cookie = cookies[i]; //得到某下标的cookies数组 // if (cookie.substring(0, cookieName.length + 2).trim() == cookieName.trim() + "=") {//如果存在该cookie的话就将cookie的值拿出来 // cookieValue = cookie.substring(cookieName.length + 2, cookie.length); // break // } // } // } // if (cookieValue != "" && cookieValue != null) {//如果存在指定的cookie值 // return false; // } // else { // // return true; // } // } // function ShowTwo(webUrl){ // alert("22"); // $.funkyUI({url:webUrl,css:{width:"600",height:"500"}}); // } //window.onload = pdfdownloadjudge;
OJGen> Vol.3 No.2, June 2013
Share This Article:
Cite This Paper >>

1000 sample comparison of MLPA and RT-PCR for carrier detection and diagnostic testing for Spinal Muscular Atrophy Type 1

Abstract Full-Text HTML Download Download as PDF (Size:111KB) PP. 111-114
DOI: 10.4236/ojgen.2013.32014    4,721 Downloads   7,046 Views   Citations
Author(s)    Leave a comment
Charles M. Strom, Ben Anderson, Mei Peng, Urjit Patel, Corey D. Braastad, Weimin Sun

Affiliation(s)

.
1Quest Diagnostics Nichols Institute, San Juan Capistrano, USA 2Athena Diagnostics, Worcester, USA.

ABSTRACT

Purpose: To compare the accuracy of a commercially available MLPA kit with a laboratory developed RT-PCR assay for the detection of SMN1 and SMN2 copy numbers in clinical samples. Methods: We developed and validated a laboratory developed real time PCR based test capable of detecting SMN1 and SMN2 copy numbers in individuals. We also validated an MLPA kit purchased from MRC Holland for the same purpose. We then analyzed a series of 1027 anonymized samples using both technologies. When discrepant results were obtained, each sample was re-analyzed at least twice using both platforms. Results: Five samples did not yield results in either assay. For SMN1 copy number analysis, 2 RT-PCR assays revealed indeterminant results and all 1020 other samples were concordant for SMN1 copy number. There were 9 discrepancies in SMN2 copy number determination mostly due to a variability in MLPA analysis. Conclusion: Both MLPA and RTPCR assays give a reliable estimate of SMN1 copy number and are therefore appropriate for population based carrier screening for Spinal Muscular Atrophy Type 1. The MLPA kit has a low incidence (<1%) of underestimating the SMN2 copy number by 1 copy, but this inconsistency is of little clinical significance and can be overcome by replicate testing.

KEYWORDS

Spinal Muscular Atrophy; Carrier Screening; SMN1; SMN2; MLPA

Cite this paper

Strom, C. , Anderson, B. , Peng, M. , Patel, U. , Braastad, C. and Sun, W. (2013) 1000 sample comparison of MLPA and RT-PCR for carrier detection and diagnostic testing for Spinal Muscular Atrophy Type 1. Open Journal of Genetics, 3, 111-114. doi: 10.4236/ojgen.2013.32014.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] [1] Lefebvre, S., Burglen, L., Reboullet, S., et al. (1995) Identification and characterization of a spinal muscular atrophy-determining gene. Cell, 80, 155-165. doi:10.1016/0092-8674(95)90460-3
[2] McAndrew, P.E., Parsons, D.W., Simard, L.R., et al. (1997) Identification of proximal spinal muscular atrophy carriers and patients by analysis of SMNT and SMNC gene copy number. The American Journal of Human Genetics, 60, 1411-1422. doi:10.1086/515465
[3] Mailman, M.D., Heinz, J.W., Papp, A.C., et al. (2002) Molecular analysis of spinal muscular atrophy and modification of the phenotype by SMN2. Genetics in Medicine, 4, 20-26. doi:10.1097/00125817-200201000-00004
[4] Ogino, S. and Wilson, R.B. (2002) Genetic testing and risk assessment for spinal muscular atrophy (SMA). Human Genetics, 111, 477-500. doi:10.1007/s00439-002-0828-x
[5] Pearn, J. (1978) Incidence, prevalence, and gene frequency studies of chronic childhood spinal muscular atrophy. Journal of Medical Geneticst, 15, 409-413. doi:10.1136/jmg.15.6.409
[6] Wirth, B., Rudnik-Schoneborn, S., Hahnen, E., Rohrig, D. and Zerres, K. (1995) Prenatal prediction in families with autosomal recessive proximal spinal muscular atrophy (5q11.2-q13.3): Molecular genetics and clinical experience in 109 cases. Prenatal Diagnosis, 15, 407-417. doi:10.1002/pd.1970150503
[7] Prior, T.W. (2008) Carrier screening for Spinal Muscular Atrophy. Genetics in Medicine, 10, 840-842. doi:10.1097/GIM.0b013e318188d069
[8] Prior, T.W., Nagan, N., Sugarman, E.A., Batish, S.D. and Braastad, C. (2011) Technical standards and guidelines for spinal muscular atrophy testing. Genetics in Medicine, 13, 686-694. doi:10.1097/GIM.0b013e318220d523
[9] (1 September 2012) http://www.mlpa.com/
[10] Gomes-Curet, I., Robinson, K.G., Funanage, V.L., Crawford, T.O., Scavina, M. and Wang, W. (2007) Robust quantification of the SMN gene copy number by realtime Taqman PCR. Neurogenetics, 8, 271-278. doi:10.1007/s10048-007-0093-1
[11] French, C., Li, C., Strom, C. and Sun, W. (2004) Detection of factor V Leiden mutation by a modified photo cross-linking oligonucleotide hybridization assay. Clinical Chemistry, 50, 296-305. doi:10.1373/clinchem.2003.023556
[12] Strom, C.M., Janeczko, R.A., Anderson, B., Redman, J., Quan, F., Buller, A., McGinniss, M. and Sun, W. (2005) Technical validation of a multiplex platform to detect 30 mutations in 8 genetic diseases prevalent in individuals of Ashkenazi Jewish descent. Genetics in Medicine, 7, 633-639. doi:10.1097/01.gim.0000187120.93597.16
[13] Strom, C.M., Janeszco, R., Quan, F., Wang, S.-B., Buller, A., McGinniss, M. and Sun, W. (2006) Technical validation of a Luminex based multiplex assay for 40 mutations in the cystic fibrosis transmembrane regulator protein. Journal of Molecular Diagnostics, 8, 371-375. doi:10.2353/jmoldx.2006.050115

  
comments powered by Disqus
OJGen Subscription
E-Mail Alert
OJGen Most popular papers
Publication Ethics & OA Statement
OJGen News
Frequently Asked Questions
Recommend to Peers
Recommend to Library
Contact Us

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.