摘要
目的建立沙门菌的聚合酶链反应-高分辨率溶解曲线(PCR-HRM)分型方法,结合DNA测序方法,比较PCR-HRM分型方法和血清学分型方法的一致性,评价该方法的临床应用价值。方法对全部32株沙门菌,先用传统的血清学分型技术进行分型,然后选择flj B、gyr B和ycf Q 3个基因为目的基因,用PCR-HRM技术对其进行分型,最后通过扩增上述3个目的基因并测序作为分型的金标准,比较血清学分型技术和PCR-HRM技术分型的一致率和正确率,分析两种方法不一致的可能原因。结果对32株所测沙门菌,血清学分型技术分出6个血清型,PCR-HRM分型技术分出5个曲线型,血清学分型技术和PCR-HRM分型技术的一致率为84.4%;以测序结果作为金标准,血清学分型技术的正确分型率为84.4%,PCR-HRM技术的正确分型率为96.9%,两种方法比较差异尚无统计学意义(P=0.156),但在本研究中PCR-HRM技术具有更高的分型正确率。结论 PCR-HRM分型方法具有简单、快速、灵敏度高和正确率高的优点,与传统的血清学分型方法具有良好的一致性,在建立标准操作规程之后,PCR-HRM技术在沙门菌分型中具有良好的应用价值。
Objective To establish a polymerase chain reaction-high resolution melting curve （PCR-HRM） typing method for Salmonella, and to compare the consistency of PCR-HRM typing method and serological typing method combined with DNA sequencing method, then evaluate the clinical applying value of the method. Methods Total of 32 cases of Salmonella, typed by traditional serotyping method firstly, then selected fljB, gyrB and ycfQ as the target genes and typed by PCR-HRM technique, the three target genes were finally amplified and sequenced as gold standard. The consistent rate and correct rate of serotyping method and PCR-HRM technique were compared and the possible inconsistent causes of the two methods were analyzed. Results For 32 cases of tested Salmonella, 6 serotypes were separated by serotyping method and 5 curve types were separated by PCR-HRM typing method. The consistency of serotyping method and PCR-HRM typing method was 84.4%. After compared with the sequencing results, the correct typing rate of serotyping method and PCR-HRM typing method were 84.4% and 96.9%, respectively, with no significant differences （P = 0.156）, but PCR-HRM typing method had a higher typing accuracy in this study. Conclusions The simple, fast, sensitive and exact PCR-HRM technique had good consistency with traditional serotyping method in Salmonella typing, after established standard operating procedures （SOP）, PCR-HRM technique had a good applying value in Salmonella typing.
出处
《中华实验和临床感染病杂志（电子版）》
CAS
2015年第3期107-110,共4页
Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)
基金
辽宁省科技攻关计划(No.2011225021)
关键词
聚合酶链反应
高分辨率溶解曲线
沙门菌
分型
Polymerase chain reaction
High-resolution melting-curve
Salmonella
Typing
