Proceeding2562

1533 การประชุมวิชาการระดับชาติมหาวิทยาลัยทักษิณ ครั้งที่ 29 ประจ�ำปี 2562 วิจัยและนวัตกรรมเพื่อการพัฒนาที่ยั่งยืน Effects of citrate phosphate dextrose adenine 1 (CPDA1) on blood group genotyping by PCR-SSP Natda Tonwong 1* , Natthaphon Na nakorn 1 , Kanyarat Saetiow 2 , Tidarat Jitmun 2 and Supattra ​Mitundee 3 Abstract Introduction : Serological technique is standard method for detection of blood group in blood bank, however, molecular technique is needed to confirm some particular cases. Source of DNA for PCR amplification is mostly from EDTA blood, in addition, CPDA1 is also commonly used in blood bank laboratory. Objective : This study aimed to investigate the effect of CPDA1 on yield of DNA at various storage times and on DNA amplification by sequence specific primer PCR (PCR-SSP). Methods : Forty of CPDA1 blood samples was stored at 4°C for 3, 10 and 30 days after venipuncture, DNA was extracted from buffy coat, measured, and amplified by PCR-SSP. Meanwhile, EDTA blood sample from the same donors was performed parallelly. Results : The highest yield of DNA from EDTA blood was retrieved from day 10, and higher than the yield from CPDA1 blood (p=0.045). On the other hand, the highest yield of DNA from CPDA1 blood was found on day 30 and was significantly higher than the yield from day 3 (p=0.004) and day 10 (p=0.011). Interestingly, on day 30, DNA yield from CPDA1 was significant higher than the yield from EDTA blood (p = 0.037). In addition, DNA from both EDTA and CPDA1 blood was successfully amplified to detect RhD blood group using PCR-SSP technique. Conclusion : These results suggested that CPDA1 can be used as anticoagulant for molecular studies without any interference. Keywords : EDTA, CPDA1, DNA extraction, PCR-SSP