摘要
目的:探索早期诊断HBV和HCV的原理和新技术.方法:主要通过PCR反应条件的探索及改变PCR反应过程中Mg2+浓度、退火温度和改用不同的Taq DNA聚合酶,研究了HBV毒株前C区基因和HCV毒株C区核心蛋白.结果:在实验选择范围内,检测HBV病毒时,4.0 mmol/L为最佳Mg2+浓度,61℃为最佳退火温度,天源公司的Taq DNA聚合酶灵敏性最高,对质粒浓度的要求达到10-9.当检测HCV病毒时,1.5 mmol/L为最佳Mg2+浓度,59.7℃为最佳退火温度,天为时代公司的iTaq聚合酶灵敏性最高,对质粒浓度的要求达到10-4.
The work aims is to devise method for early diagnosis of HBV and HCV. The constructed plasmid （pGEM-T-HBV （C region/C gene） and pGEM-T-HCV （core）） were used as the template. Optimization of PCR conditions was made by changing Mg^2＋ concentration, annealing temperature and different Taq DNA polymerase. The results show that the optimal conditions of HBV virus detection is 4. 0 mmol/L as the best Mg^2＋ concentration, 61 ℃ as the best annealing temperature and the Taq DNA polymerase from Tian yuan company having the highest sensitivity and specificity. The optimal plasmid concentration was of 10^-9. While the optimal conditions of HCV virus detection is 1.5 mmol/L as the best Mg^2- concentration, 59.7 ℃ as the best annealing temperature and the Tianweishidai company＇s Taq DNA polymerase has good in the sensitivity and specificity. The optimal plasmid concentration was of 10^-4.
出处
《中南民族大学学报（自然科学版）》
CAS
2008年第3期36-39,共4页
Journal of South-Central University for Nationalities：Natural Science Edition
基金
澳大利亚国家自然科学基金资助项目(ARClargegrant(A)0087128)
