Balkan Journal of Medical Genetics

ANALYSIS OF HUMAN BRADYKININ RECEPTOR GENE AND ENDOTHELIAL NITRIC OXIDE SYNTHASE GENE POLYMORPHISMS IN END-STAGE RENAL DISEASE AMONG MALAYSIANS
Vasudevan R, Ismail P, Jaafar NI, Mohamad NA, Etemad E, Wan Aliaa WS, Eshkor S a These authors contributed equally to this article.
*Corresponding Author: Professor Dr. Patimah Ismail, Department of Biomedical Sciences, Faculty of Medical and Health Science, Universiti Putra Malaysia, Serdang 43400, Selangor DE, Malaysia. Tel.: +60-3-8947-2314. Fax: +60-3-8943-6178. E-mail: patimashimail@gmail.com and Dr. Ramachandran Vasudevan, Institute of Gerontology, Universiti Putra Malaysia, Serdang, Selangor DE, 43400, Malaysia. Tel.: +60-3-8947-2752. Fax: +60-3-8947-2738. E-mail: vasuphd@gmail.com.
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MATERIALS AND METHODS

The study was approved by the Ethical Committee of the Faculty of Medicine and Health Science (RUGS project no. 91104) and permission was obtained from the National Kidney Foundation (NKF) of Malaysia. A total of 320 subjects were approached and 300 [150 patients from the NKF undergoing dialysis treatment and 150 unrelated healthy individuals, randomly selected by conducting health screening programs at various places in and around the Universiti Putra Malaysia (UPM) Serdang, Malaysia area], were recruited for this study; some subjects were excluded due to inconsistent genotyping results and extreme values. Written consent was obtained from all subjects who participated in this study and buccal cells were collected from ESRD subjects using cytology brushes. Genomic DNA was extracted to determine the genetic polymorphisms of the eNOS and B2R genes. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was carried out for all subjects according to the available protocols [8,9]. All the amplified products were separated using 2.0% agarose gel electrophoresis and the gel was stained using Gel Red (Biotium, Hayward, CA, USA). The stained gel was visualized under UV light using an Alpha Imager (Alpha Innotech, San Leandro, CA, USA). Identical results were obtained when genotyping was performed on two separate occasions for 10.0% of the samples. All the statistical analyses were carried out using the Statistical Package for the Statistical Sciences (SPSS) (Chicago, IL, USA) software, version 18.0 for Microsoft Windows. A p value of <0.05 was considered to be significant.

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