CAJ | 학술논문

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fljB、gyrB和ycfQ 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。
목적：건립사문균적취합매련반응-고분변솔용해곡선（PCR-HRM）분형방법，결합DNA측서방법，비교PCR-HRM분형방법화혈청학분형방법적일치성，평개해방법적림상응용개치。방법대전부32주사문균，선용전통적혈청학분형기술진행분형，연후선택fljB、gyrB화ycfQ 3개기인위목적기인，용PCR-HRM기술대기진행분형，최후통과확증상술3개목적기인병측서작위분형적금표준，비교혈청학분형기술화PCR-HRM기술분형적일치솔화정학솔，분석량충방법불일치적가능원인。결과대32주소측사문균，혈청학분형기술분출6개혈청형，PCR-HRM분형기술분출5개곡선형，혈청학분형기술화PCR-HRM분형기술적일치솔위84.4%；이측서결과작위금표준，혈청학분형기술적정학분형솔위84.4%，PCR-HRM기술적정학분형솔위96.9%，량충방법비교차이상무통계학의의（P=0.156），단재본연구중PCR-HRM기술구유경고적분형정학솔。결론 PCR-HRM분형방법구유간단、쾌속、령민도고화정학솔고적우점，여전통적혈청학분형방법구유량호적일치성，재건립표준조작규정지후，PCR-HRM기술재사문균분형중구유량호적응용개치。
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 ifrstly, then selected lfjB, gyrB and ycfQ as the target genes and typed by PCR-HRM technique, the three target genes were ifnally ampliifed 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 signiifcant 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.