检验医学与临床2018，Vol.15Issue(6)：737-739,743,4.DOI:10.3969/j.issn.1672-9455.2018.06.001
实时定量PCR检测流感嗜血杆菌感染实验室诊断方法的建立
Laboratory diagnostic method establishment of real-time quantitative PCR for detecting Haemophilus influenza infection
摘要
Abstract
Objective To establish a laboratory diagnostic method of SYBR Green dye method of real-time quantitative PCR for detecting Haemophilus influenza(Hi)infection.Methods T he primers aiming at L-fucu-lokinase(fucK)gene were designed and synthesized.The Hi fucK gene was amplified,the plasmid was con-structed,after 10 fold gradient dilution,the standard curve was drawn with 10 -108copy of reaction as the standard substance,the sensitivity of newly established laboratory diagnostic method was tested;the DNA samples of Mycoplasma pneumoniae standard strain,Mycoplasma hominis standard strain,Enterobacter cloa-cae standard strain,Staphylococcus aureus standard strain,Klebsiella pneumoniae standard strain and Esche-richia coli standard strain preserved in the laboratory were adopted for detecting the specificity of newly estab-lished laboratory diagnostic method;then the newly established laboratory diagnostic method was used in the detection of clinical samples.Results This newly established laboratory diagnostic method had higher sensi-tivity,its detectable copy number was 10 copy Hi DNA;this method had higher specificity and could success-fully distinguish several pathogenic DNA;in the application of this method for 226 clinical throat swab sam-ples,92 cases of Hi infection were detected out with the positive rate of 40.70%.Conclusion The real-time quantitative PCR for detecting Hi infection established by this study has higher sensitivity and higher specific-ity,is easy to operate,uses short time and can be used in the laboratory diagnosis of clinical Hi infection.关键词
流感嗜血杆菌/儿童/呼吸道感染/实时定量聚合酶链反应Key words
Haemophilus influenzae/children/respiratory tract infections/real time quantitative PCR分类
医药卫生引用本文复制引用
蔚然,王良玉,高琦,胡文娟,窦海伟,向莉,辛德莉,郭东星..实时定量PCR检测流感嗜血杆菌感染实验室诊断方法的建立[J].检验医学与临床,2018,15(6):737-739,743,4.基金项目
首都市民健康培育项目(Z161100000116088、Z131100006813044). （Z161100000116088、Z131100006813044）
