J Clin Lab Anal. 2018;32:e22300. wileyonlinelibrary.com/journal/jcla
1 of 6
© 2017 Wiley Periodicals, Inc.
Measurement of absolute copy number variation of
Glutathione S- Transferase M1 gene by digital droplet PCR
and association analysis in Tunisian Rheumatoid Arthritis
| Mohamed Sahbi Ben Kilani
| Mariem Ben Hamad
| Nadia Mahfoudh
| Zouheir Bahloul
| Leila Keskes
| Abdellatif Maalej
Laboratory of Human Molecular
Genetics, Faculty of Medecine, Sfax, Tunisia
Hospital Hedi Chaker, Sfax, Tunisia
Chaker, Sfax, Tunisia
Faculty of Medicine, Sfax, Tunisia.
Ministry of Higher Education and Scientific
Research of the republic of Tunisia
Background: The investigation of copy number variations (CNVs) analysis of candidate
genes is currently an important research area in modulating human diseases. We
aimed to quantify CNVs in glutathione S- transferase M1 (GSTM1) gene and determine
an innovative technique for quantification.
Using a recently powerful approach of digital droplet PCR (ddPCR), we quantified
GSTM1 gene to determine the presence of no, one, or multiple copy number (CN) at
high levels of sensitivity and specificity. Odds ratio and Fisher exact test were per-
formed to estimate the association risk for GSTM1CNVsinRA.
copy) of GSTM1 was found to be a significant risk factor for anti- cyclic citrullinated
tion of CNVs and suggests that the variation in the CN of GSTM1 is associated with
bility to the disease in our Tunisian sample.
copynumbervariants,dropletdigitalPCR,GSTM1, rheumatoid arthritis
1 | INTRODUCTION
disease affecting approximately 1% of the adult population world-
wide. It is characterized by the symmetrical inflammation of the sy-
novial joints, increased cytokine production and pannus development,
leading, in fact, to a progressive destruction of the cartilage and bones.
lation and family studies have indicated that genetic factors are major
which are estimated to be as high
class II genes are depicted as being the most potent factor associated