Simultaneous detection of 15 respiratory pathogens with a fluorescence probe melting curve analysis-based multiplex real-time PCR assay.

International journal of molecular epidemiology and genetics Pub Date : 2019-04-15 eCollection Date: 2019-01-01
Shengyun Liao, Lingli Wang, Xiang Ji, Jiandong Chen, Qiang Li, Lan Ma
{"title":"Simultaneous detection of 15 respiratory pathogens with a fluorescence probe melting curve analysis-based multiplex real-time PCR assay.","authors":"Shengyun Liao,&nbsp;Lingli Wang,&nbsp;Xiang Ji,&nbsp;Jiandong Chen,&nbsp;Qiang Li,&nbsp;Lan Ma","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Acute respiratory tract infections are common worldwide and caused by a great diversity of pathogens. A rapid and accurate diagnosis method of respiratory infection is crucial for timely clinical intervention. Here, by combining fluorescence melting curve analysis and multiplex real-time assay, we developed a novel method which can simultaneously detect 15 respiratory viruses. The specificity for target genes was 100%, as assessed with a panel of 47 respiratory pathogens, which indicated no cross-reactions. The assay's limits of detection at the nucleic acid level ranged from 5 copies/μL to 500 copies/μL nucleic acids. Compared with conventional culture method, our assay showed more than 75% sensitivity and 100% specificity for each respiratory pathogen in 384 clinical samples. Even more, the kappa correlation for all the pathogens ranged from 0.86 to 1.00. Overall, this method has the characteristics of high throughput, low cost and high sensitivity and precision, which demonstrated our method is well suited for routine clinical testing in respiratory infection.</p>","PeriodicalId":73460,"journal":{"name":"International journal of molecular epidemiology and genetics","volume":"10 2","pages":"29-37"},"PeriodicalIF":0.0000,"publicationDate":"2019-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6526376/pdf/ijmeg0010-0029.pdf","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"International journal of molecular epidemiology and genetics","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2019/1/1 0:00:00","PubModel":"eCollection","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Acute respiratory tract infections are common worldwide and caused by a great diversity of pathogens. A rapid and accurate diagnosis method of respiratory infection is crucial for timely clinical intervention. Here, by combining fluorescence melting curve analysis and multiplex real-time assay, we developed a novel method which can simultaneously detect 15 respiratory viruses. The specificity for target genes was 100%, as assessed with a panel of 47 respiratory pathogens, which indicated no cross-reactions. The assay's limits of detection at the nucleic acid level ranged from 5 copies/μL to 500 copies/μL nucleic acids. Compared with conventional culture method, our assay showed more than 75% sensitivity and 100% specificity for each respiratory pathogen in 384 clinical samples. Even more, the kappa correlation for all the pathogens ranged from 0.86 to 1.00. Overall, this method has the characteristics of high throughput, low cost and high sensitivity and precision, which demonstrated our method is well suited for routine clinical testing in respiratory infection.

Abstract Image

Abstract Image

分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
基于荧光探针熔融曲线分析的多重实时荧光定量PCR同时检测15种呼吸道病原体。
急性呼吸道感染在世界范围内很常见,由多种病原体引起。快速准确的诊断方法对临床及时干预至关重要。本文将荧光熔化曲线分析与多重实时检测相结合,建立了一种可同时检测15种呼吸道病毒的新方法。靶基因的特异性为100%,由47种呼吸道病原体组成的小组进行评估,表明无交叉反应。该方法在核酸水平上的检测限为5拷贝/μL ~ 500拷贝/μL。与常规培养法相比,384份临床标本中各呼吸道病原菌的检测灵敏度均在75%以上,特异性为100%。更重要的是,所有病原体的kappa相关性在0.86到1.00之间。总体而言,该方法具有高通量、低成本、高灵敏度和精密度的特点,适用于呼吸道感染的常规临床检测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Exploring the effects of homocysteine metabolism in osteoporosis management in Indian adult females. P. falciparum genetic markers associated with drug resistance from patients with treatment failure in the Southern part of Senegal in 2017. Abundance and diversity of methicillin-resistant bacteria from bathroom surfaces at workplaces using CHROMagar media, 16S, and dnaJ gene sequence typing. Analysis of carbapenem-resistant Acinetobacter baumannii carbapenemase gene distribution and biofilm formation. Facial and ocular manifestations of male patients affected by the HUWE1-related intellectual developmental disorder.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1