RRML - Sanger sequencing of MMR genes in a one-plate system
AMLR

ISSN online: 2284-5623

ISSN-L: 1841-6624

Rejection rate (2016): 55%

Română English


SCImago Journal & Country Rank


Advanced search


Top 10 downloaded articles
- July 2018 -
 
Serum hs-CRP, YKL-40 and brain... 14
Non-invasive quantification of... 11
Postoperative interleukin-8 le... 10
Candida albicans isolates from... 9
Rationale and Design of a Pros... 9
Can laboratory assays for meas... 8
Phenotypic and molecular ident... 7
Involvement of inflammatory cy... 7
Arylesterase activity of Parao... 7
Ischemic modified albumin incr... 6

Log in

Concept, Design & Programming
Dr. Adrian Man

   
 
Nr. 26(2)/2018 DOI:10.2478/rrlm-2018-0008
XML
TXT

Research article

Sanger sequencing of MMR genes in a one-plate system

Lucian Negura, Anca Negura

Correspondence should be addressed to: Lucian Negura

Abstract:

Both incidence and mortality of colorectal cancer (CRC) in Romania have shown a continuous increase during the last decades. Hereditary Non-Polyposic Colorectal Cancer (HNPCC), also known as Lynch syndrome, is mainly attributable to mismatch repair (MMR) genes MSH2, MSH6, and MLH1. Individuals carrying germ-line mutations of these genes present high lifetime risk of colorectal and other cancers, compared to non-carriers. Oncogenetics is developed worldwide nowadays, for identifying hereditary predisposition to cancer and offering appropriate clinical follow-up to patients and mutation carriers in Lynch families. Molecular oncogenetic diagnosis in Lynch syndrome is based on complete Sanger sequencing of entire MMR genes, which is time and resources consuming, therefore needing an appropriate and adapted optimization. Conventional sequencing requires a sufficient number of available samples to be processed simultaneously, which increases the waiting time for diagnostic results. Complete analysis for only one patient meets difficult technical problems due to the complex co-amplification of all gene regions of interest within the same conditions, therefore increasing the costs and reducing the cost-effectiveness of the test. Here we present an original and robust technical protocol for sequencing the entire MSH2, MSH6, and MLH1 coding sequence for one patient in a single PCR plate. Our optimized and verified system overcomes all technical problems and offers a quick, robust, and cost-effective possibility to personalize molecular oncogenetic diagnosis in Lynch syndrome.

Keywords: Lynch syndrome; MMR genes; Sanger sequencing; molecular diagnostic; cost-effectiveness

Received: 21.8.2017
Accepted: 10.1.2018
Published: 5.2.2018

 
  PDF Download full text PDF
(1174 KB)
     
 
How to cite
Negura L, Negura A. Sanger sequencing of MMR genes in a one-plate system. Rev Romana Med Lab. 2018;26(2):153-63. DOI:10.2478/rrlm-2018-0008