摘要
目的建立一种高分辨率熔解曲线(HRM)-非标记探针的方法用于检测CCL2-2518T>C的单核苷酸多态性(SNP)。方法针对CCL2-2518 T>C,分别设计出针对野生型和突变型的两个非标记探针,采用HRM-非标记探针法对71名健康人群CCL2-2518位点的SNP进行检测,并采用TaqMan分型法进行验证。结果 HRM-非标记法的两个探针与TaqMan分型法分型结果一致,准确率100%;针对突变型(C)的探针明显好于针对野生型(T)的探针,其检测野生纯合型和突变纯合型的探针峰的温度之差分别为5.5℃和2.1℃。结论本研究建立的CCL2-2518 T>C位点突变检测的HRM-非标记探针方法是一种简单、廉价、灵敏、准确和快速的技术,适用于临床检测。
ObjectiveTo establish a high resolution melting analysis with unlabeled probes for genotyping CCL2-2518 T〉C.Methods Two unlabeled probes were designed. One was complementary to wild type and another was complementary to the mutant type. 71 health people were performed by HRM with unlabeled probes for genotyping CCL2-2518 T〉C, and TaqMan method was used to verify the results from above.Results The results using the HRM with unlabeled probes were consistent with that using TaqMan method. The difference between the temperature in two probe peaks by using the probe complementary to wild type was 2.1℃; and the difference between the temperature in two probe peaks by using the probe complementary to mutant type was 5.5℃.Conclusion Establishment the high resolution melting analysis with unlabeled probes for genotyping CCL2-2518 T〉C is a simple, economic, sensitive, accurate, and fast technique, which could be used in clinical inspection.
出处
《中华临床医师杂志（电子版）》
CAS
2014年第13期92-94,共3页
Chinese Journal of Clinicians(Electronic Edition)
基金
中国博士后科学基金(2011M500206)
