连接酶检测反应与实时荧光定量聚合酶链反应检测慢性乙型肝炎患者低丰度YMDD变异体的比较
Comparison of ligase detection reaction and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B.
作者信息
Wang Xiao-Ling, Xie Song-Gang, Zhang Ling, Yang Wei-Xia, Wang Xing, Jin Hong-Zhi
机构信息
Laboratory Center, Subei People's Hospital, 98 West Nantong Road, Yangzhou 225001, Jiangsu Province, China.
出版信息
World J Gastroenterol. 2008 Jan 7;14(1):120-4. doi: 10.3748/wjg.14.120.
AIM
To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.
METHODS
Mixtures of plasmids and serum samples from 52 chronic hepatitis B patients with low abundant lamivudine-resistant mutations were tested with LDR and real-time PCR. Time required and reagent cost for both assays were evaluated.
RESULTS
Real-time PCR detected 100, 50, 10, 1 and 0.1% of YIDD plasmid, whereas LDR detected 100, 50, 10, 1, 0.1, and 0.01% of YIDD plasmid, in mixtures with YMDD plasmid of 10(6) copies/mL. Among the 52 clinical serum samples, completely concordant results were obtained for all samples by both assays, and 39 YIDD, 9 YVDD, and 4 YIDD/YVDD were detected. Cost and time required for LDR and real-time PCR are 60/80 CNY (8/10.7 US dollars) and 4.5/2.5 h, respectively.
CONCLUSION
LDR and real-time PCR are both sensitive and inexpensive methods for monitoring low abundant YMDD mutants during lamivudine therapy in patients with chronic hepatitis B. LDR is more sensitive and less expensive, while real-time PCR is more rapid.
目的
比较连接酶检测反应（LDR）和实时荧光定量聚合酶链反应（real-time PCR）检测慢性乙型肝炎感染患者中低丰度YMDD突变体的效果。
方法
采用LDR和实时荧光定量聚合酶链反应检测52例慢性乙型肝炎患者低丰度拉米夫定耐药突变的质粒和血清样本混合物。评估两种检测方法所需时间和试剂成本。
结果
在与10(6)拷贝/mL YMDD质粒混合的样本中，实时荧光定量聚合酶链反应能检测出100%、50%、10%、1%和0.1%的YIDD质粒，而LDR能检测出100%、50%、10%、1%、0.1%和0.01%的YIDD质粒。在52份临床血清样本中，两种检测方法对所有样本的检测结果完全一致，共检测出39例YIDD、9例YVDD和4例YIDD/YVDD。LDR和实时荧光定量聚合酶链反应的成本分别为60/80元人民币（8/10.7美元），所需时间分别为4.5/2.5小时。
结论
LDR和实时荧光定量聚合酶链反应都是监测慢性乙型肝炎患者拉米夫定治疗期间低丰度YMDD突变体的敏感且廉价的方法。LDR更敏感且成本更低，而实时荧光定量聚合酶链反应更快速。
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