Open AccessMeasurement of absolute copy number variation of Glutathione S‐Transferase M1 gene by digital droplet PCR and association analysis in Tunisian Rheumatoid Arthritis population
Author(s) -
Achour Yosser,
Ben Kilani Mohamed Sahbi,
Ben Hamad Mariem,
Marzouk Sameh,
Mahfoudh Nadia,
Bahloul Zouheir,
Keskes Leila,
PetitTeixeira Elisabeth,
Maalej Abdellatif
Publication year - 2018
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22300
Subject(s) - copy number variation , rheumatoid arthritis , glutathione s transferase , gene , genetics , digital polymerase chain reaction , real time polymerase chain reaction , biology , population , odds ratio , immunology , medicine , genome , polymerase chain reaction , glutathione , enzyme , biochemistry , environmental health
Background The investigation of copy number variations ( CNV s) analysis of candidate genes is currently an important research area in modulating human diseases. We aimed to quantify CNV s in glutathione S‐transferase M1 ( GSTM 1 ) gene and determine its genetic contribution in Tunisian rheumatoid arthritis ( RA ) and its subsets through an innovative technique for quantification. Methods A total of 165 RA cases and 102 healthy controls were included in the study. Using a recently powerful approach of digital droplet PCR (dd PCR ), we quantified GSTM 1 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 performed to estimate the association risk for GSTM 1 CNV s in RA . Results Copy number identified by dd PCR was 0, 1, and 2 copies per diploid genome. A high frequency of ‘0’ copy was revealed with 54% in RA patients. The deletion (‘0’ copy) of GSTM 1 was found to be a significant risk factor for anti‐cyclic citrullinated peptide (anti‐ CCP ) positive RA ( OR =4.16, CI 95% =[1.17‐14.7]). In addition, a lack of association was found when comparing between the CNV s of RA patients and those of controls. Conclusion This study highlights the powerful accuracy of dd PCR for the quantification of CNV s and suggests that the variation in the CN of GSTM 1 is associated with anti‐ CCP positivity in RA . However, it does not indicate a specific role in the susceptibility to the disease in our Tunisian sample.