NDM1基因常规PCR与荧光定量PCR检测方法的建立及其检测结果比较

doi:10.7694/jldxyxb20130342

吉林大学学报(医学版) ›› 2013, Vol. 39 ›› Issue (3): 625-629.doi: 10.7694/jldxyxb20130342

NDM1基因常规PCR与荧光定量PCR检测方法的建立及其检测结果比较

陈宣男¹，全首祯^2

1.中国医学科学院阜外心血管病医院临床检验中心，北京 100037;2.空军总医院临床检验中心，北京 100142

收稿日期:2012-12-27 出版日期:2013-05-28 发布日期:2013-07-01
作者简介:陈宣男（1984-），女，吉林省通化市人，研究实习员，医学硕士，主要从事院内感染细菌和血小板代谢标志物的相关研究。
基金资助:
中国医学科学院青年基金资助课题（2011-F01）

Establishment of conventional PCR and fluorescence quantitative PCR method in detection of NDM1 gene andcomparison of detection results

CHEN Xuan-nan¹,QUAN Shou-zhen²

1.Clinical Inspection Center,Fuwai Cardiovascular Disease Hospital,Chinese Academy of Medical Sciences,Beijing 100037,China;2.Clinical Inspection Center,Air Force General Hospital,Beijing 100142,China

Received:2012-12-27 Online:2013-05-28 Published:2013-07-01

摘要/Abstract

摘要： 目的：建立NDM1基因常规PCR和荧光定量PCR（FQ-PCR）检测方法，并对其检测结果进行比较。方法：设计3对NDM1基因常规PCR与FQ-PCR引物，建立常规PCR与FQ-PCR反应体系和条件，比较2种检测方法的特异性、灵敏度和对模拟临床样品的检测效果。结果：成功建立NDM1基因常规PCR和FQ-PCR检测方法；特异性检测,常规PCR与FQ-PCR检测7种常见病原菌均为阴性；灵敏度检测,常规PCR检测限为10⁶mL^-1 ，FQ-PCR检测限为10⁴mL^-1；模拟临床样品检测，常规PCR和FQ-PCR检测结果一致。结论：常规PCR与FQ-PCR均有很好的特异性，FQ-PCR灵敏度比常规PCR高100倍。

关键词: 新德里金属-&beta, -内酰胺酶1基因, 常规PCR, 荧光定量PCR

Abstract: Abstract:Objective To establish conventional PCR and fluorescence quantitative PCR(FQ-PCR) methods for detection of NDM1 gene and to compare the detection results of the two methods.Methods Three pairs of primers that specifically recognized the NDM1 gene were respectively designed,and their reaction systems and parameters were confirmed.Under the optimized conditions,the conventional PCR and FQ-PCR methods were evaluated for the specificity,sensitivity and detection effects on simulative clinical samples.Results Conventional PCR and FQ-PCR methods were successfully established.The results of specificity analysis showed that conventional PCR and FQ-PCR both presented negative resuts for seven familiar pathogenic bacteria.The threshold of FQ-PCR was 104 mL-1 and that of conventional PCR was 10⁶ mL^-1.The detection results of s
imulative clinical samples indicated that they had accordant results.Conclusion Both conventional PCR and FQ-PCR methods possess good specificity,and the sensitivity of FQ-PCR is 100 times higher than conventional PCR.

Key words: new delhi metallo-&beta, -lactamas 1 gene, conventional PCR, fluorescence quantitative-PCR

中图分类号:

陈宣男，全首祯. NDM1基因常规PCR与荧光定量PCR检测方法的建立及其检测结果比较[J]. 吉林大学学报(医学版), 2013, 39(3): 625-629.

CHEN Xuan-nan,QUAN Shou-zhen. Establishment of conventional PCR and fluorescence quantitative PCR method in detection of NDM1 gene andcomparison of detection results[J]. Journal of Jilin University Medicine Edition, 2013, 39(3): 625-629.

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NDM1基因常规PCR与荧光定量PCR检测方法的建立及其检测结果比较

Establishment of conventional PCR and fluorescence quantitative PCR method in detection of NDM1 gene andcomparison of detection results

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