Open Access Study protocol

Identification of genetic contribution to ischemic stroke by screening of single nucleotide polymorphisms in stroke patients by using a case control study design

Amit Kumar1, Ram Sagar1, Pradeep Kumar1, Jitendra K Sahu2, Ashoo Grover3, Achal K Srivastava1, S Vivekanandhan4 and Kameshwar Prasad1*

Author Affiliations

1 Department of Neurology, Room No. 704, Neurosciences Centre, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India

2 Division of Pediatric Neurology, Department of Pediatrics, PGIMER, Chandigarh, India

3 Indian Council of Medical Research, New Delhi, India

4 Department of Neurobiochemistry, All India Institute of Medical Sciences, New Delhi, India

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BMC Neurology 2013, 13:136  doi:10.1186/1471-2377-13-136

Published: 3 October 2013



Stroke is the second most common cause of death and disability worldwide. It is a multi-factorial disease influenced by both environmental and genetic factors. Studies from the different ethnic regions of world have reported variable results on association of Apolioprotein E (APOE), Methylenetetrahydrofolate reductase (MTHFR), Endothelial Nitric Oxide Synthase (ENOS), Factor V Leiden (F5), Cytochrome P450 4F2 (CYP4F2), beta-fibrinogen and Phosphodiesterase 4D (PDE4D) gene in stroke. There has been substantial evidence from the European descent genetic studies showing that genetic risk of stroke varies as per specific subtypes of ischemic stroke.

This study aims to test the hypothesis that above mentioned encoding gene polymorphisms are associated with stroke and to determine whether risk varies as per specific subtypes of stroke.


The study design would be case–control study. Six hundred cases with diagnosis of stroke and 600 age and sex matched controls will be recruited. Controls will be matched in 1:1 ratio. Baseline and demographic data will be collected in standardized data collection form. Four ml of blood will be collected in EDTA coated vial and will be used for DNA isolation. Genotyping will be done by using PCR-RFLP method. For the reconfirmation of RFLP results, PCR product of each genotype in triplet for all the selected polymorphism will be sent for DNA sequencing. Data will be analyzed using conditional logistic regression to determine odds ratio associated with the above genes.


This protocol will assess the association of above mentioned gene polymorphisms with ischemic stroke in North Indian Population. This study will also helpful to determine genetic component of stroke and whether variation in genetic risk as per different subtypes of stroke.