Objectives: The aim of the present study was to develop a simple, quick and cheap method to process whole-blood samples for the molecular techniques polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) without the use of expensive reagents or sophisticated machines. Materials and Methods: Venous whole-blood samples were collected from 40 individuals. The samples were frozen at –80°C, and then rapidly thawed at 37°C. Each sample was incubated with distilled water, then boiled in a microwave and centrifuged. The supernatant was taken directly for PCR and RFLP. For comparison, PCR and RFLP were performed on DNA purified from the same samples using the phenol-chloroform method and two commercial DNA extraction kits. Results: PCR/RFLP results using the presented method were qualitatively similar to those obtained by DNA extracted using the other three methods. Conclusion: The presented method proved to be a simpler and cheaper way of processing whole-blood samples for PCR and RFLP analyses.

Factor V Leiden (FVL) mutation (G1691A) is a risk factor for development of venous thromboembolic disorders. FVL was found mostly in Caucasians (1-15%) but was almost absent in non-Caucasians. Studies on Arab patients and populations revealed very inconsistent results. This study reports FVL in Arabs living in Kuwait with a focus on the nationality of the Arab subjects studied. Whole-blood samples were collected from 400 healthy Arabs who were 268 Kuwaitis (67%), 50 Syrians (12.5%), 34 Jordanians (8.5%), 8 Palestinians (2%) and 40 Egyptians (10%). DNA extraction was carried out for these blood samples and real-time PCR was performed to detect the presence of FVL. Generally, 36 cases (9%) had the mutation (33 were heterozygous and 3 were homozygous), with an allelic frequency of 0.049. The prevalence of FVL differed in different Arabic cases: Kuwaitis 4.5%, Egyptians 15%, Syrians 16%, Jordanians 23.5% and Palestinians 25%. The allelic frequency was 0.022 in the Kuwaitis and 0.088-0.132 in non-Kuwaitis. The three homozygous cases were from Syria, Jordan and Egypt. In conclusion, the prevalence of FVL in Arabs living in Kuwait is as high as in Caucasians. There is a difference in prevalence among Arabs themselves, being relatively lower in Kuwaitis than in non-Kuwaitis. Keywords: Arabs; factor V Leiden mutation; Kuwait; real-time PCR INTRODUCTIONIn 1994, a point mutation (G1691A) in the gene encoding for the human clotting cofactor V (FV) was discovered and called factor V Leiden mutation (FVL) after the Dutch city 'Leiden' where it was discovered. 1 In consequence to this genetic mutation, the translated FV molecule has an 'arginine' amino acid in place of the normal 'glutamine' at the amino-acid residue number 506 of the molecule (Arg506Gln). 1,2 This apparently simple point mutation was found to lead to a condition called activated protein C resistance, which causes, in carriers of the mutation, a serious clinical condition known as 'hypercoagulability' or 'thrombophilia' , which is very often manifested clinically as venous thromboembolic disorders (VTE). 3-6 VTE is a significant cause of morbidity and mortality in many countries, including Kuwait, with an annual incidence of 1 out of 1000. 3,4,7,8 Studies have shown that people with FVL had a higher risk of developing VTE (10-fold and 140-fold increased risk in heterozygous and homozygous carriers, respectively). 1,2,5,6,[9][10][11][12] Several studies were conducted worldwide to determine the prevalence and risk of FVL in different countries and ethnic groups. FVL was reported to be highly present in populations of Caucasian origin living in Europe, United States and Australia, with a prevalence of 15-65% among VTE patients and 1-15% in the normal populations studied. 1,4,6,7,[10][11][12][13][14][15][16][17][18][19][20] On the other hand, FVL was almost absent in other populations such as Africans, South-East Asians, Chinese, Japanese, American Indians, Greenland Eskimos and Aboriginals of Australia. 7,17,[20][21][22][23][24][25] In Kuwait, VTE is quite commo...

The prevalence of APC-R and FVL is quite high in Arabs living in Kuwait, being comparable with the prevalence reported in Whites, although being toward the lowest values reported there.

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Objective: It was the aim of this study to report a new point mutation in the clotting factor V gene in the general Arab population. Subjects and Methods: The HR2 haplotype was tested in 288 Arabs living in Kuwait – 188 patients with venous thromboembolic disorders (VTE) and 100 healthy subjects – using polymerase chain reaction and restriction fragment length polymorphism techniques. The presence of the new mutation was verified by DNA sequencing. Results: Two (1.06%) VTE patients had guanine instead of the wild-type adenine at nucleotide number 3935 (A3935G) of the factor V gene. This mutation caused a histidine to arginine change in amino acid number 1254 of the factor V molecule. The new mutation is termed ‘factor V Kuwait’ (His1254Arg) and was absent in the 100 healthy subjects. Conclusion: It appears that factor V Kuwait could be a risk factor for developing VTE in Arabs. A larger study is needed to confirm this observation.