• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

基于SYBR Green I荧光染料的实时荧光定量PCR技术在生物样本中菌株检测的评估

Evaluation of Real-time PCR Based on SYBR Green I Fluorescent Dye for Detection of Strains in Biological Samples.

作者信息

Kędrak-Jabłońska Agnieszka, Budniak Sylwia, Szczawińska Anna, Reksa Monika, Krupa Marek, Szulowski Krzysztof

机构信息

Department of Microbiology, National Veterinary Research Institute, 24-100 Pulawy, Poland.

出版信息

J Vet Res. 2018 Dec 31;62(4):549-554. doi: 10.2478/jvetres-2018-0075. eCollection 2018 Dec.

DOI:10.2478/jvetres-2018-0075
PMID:30729215
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6364171/
Abstract

INTRODUCTION

The aim of the study was the application and evaluation of real-time PCRs based on the fluorescence of SYBR Green I intercalating dye for the detection of three genes in contaminated liver and blood samples. The goals for detection were gene as a chromosomal marker, gene located on plasmid pXO1, and gene located on plasmid pXO2.

MATERIAL AND METHODS

Five strains were used for the experiments. Additionally, single strains of other species of the genus , . , , , and , and strains of six other species were used for evaluation of the specificity of the tests. Three SYBR Green I real-time PCRs were conducted allowing confirmation of in the biological samples.

RESULTS

The observation of amplification curves in real-time PCRs enabled the detection of the chromosomally encoded gene, gene, and gene of . The specificity of the tests was confirmed by estimation of the melting temperature of the PCR products. The sensitivity and linearity of the reactions were determined using regression coefficients. Strains of other microbial species did not reveal real-time PCR products.

CONCLUSION

All real-time PCRs for the detection of in biological samples demonstrated a significant sensitivity and high specificity.

摘要

引言

本研究的目的是应用基于SYBR Green I嵌入染料荧光的实时荧光定量PCR技术检测污染肝脏和血液样本中的三个基因。检测目标为作为染色体标记的基因、位于质粒pXO1上的基因以及位于质粒pXO2上的基因。

材料与方法

使用5株菌株进行实验。此外，还使用了属内其他物种的单株菌株，如、、、、和，以及其他六个物种的菌株来评估检测的特异性。进行了三次SYBR Green I实时荧光定量PCR，以确认生物样本中的情况。

结果

实时荧光定量PCR中扩增曲线的观察能够检测到的染色体编码基因、基因和基因。通过估计PCR产物的熔解温度来确认检测的特异性。使用回归系数确定反应的灵敏度和线性。其他微生物物种的菌株未显示实时荧光定量PCR产物。

结论

所有用于检测生物样本中情况的实时荧光定量PCR均显示出显著的灵敏度和高特异性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/967a/6364171/eaaca81fd005/jvetres-62-549-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/967a/6364171/7e207991bcc7/jvetres-62-549-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/967a/6364171/d48d0053edb4/jvetres-62-549-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/967a/6364171/eaaca81fd005/jvetres-62-549-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/967a/6364171/7e207991bcc7/jvetres-62-549-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/967a/6364171/d48d0053edb4/jvetres-62-549-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/967a/6364171/eaaca81fd005/jvetres-62-549-g003.jpg

相似文献

1
Evaluation of Real-time PCR Based on SYBR Green I Fluorescent Dye for Detection of Strains in Biological Samples.基于SYBR Green I荧光染料的实时荧光定量PCR技术在生物样本中菌株检测的评估
J Vet Res. 2018 Dec 31;62(4):549-554. doi: 10.2478/jvetres-2018-0075. eCollection 2018 Dec.
2
Detection of Spp. and in Biological Samples by SYBR Green I and TaqMan Probe-based Real-time PCRs.通过SYBR Green I和基于TaqMan探针的实时聚合酶链反应检测生物样品中的特定物种和（此处原文不完整，缺少具体内容）
J Vet Res. 2017 Dec 27;61(4):427-432. doi: 10.1515/jvetres-2017-0069. eCollection 2017 Dec.
3
Rapid genotypic detection of Bacillus anthracis and the Bacillus cereus group by multiplex real-time PCR melting curve analysis.通过多重实时PCR熔解曲线分析快速进行炭疽芽孢杆菌和蜡样芽孢杆菌群的基因分型检测。
FEMS Immunol Med Microbiol. 2005 Feb 1;43(2):301-10. doi: 10.1016/j.femsim.2004.10.005.
4
Identification and characterization of Bacillus anthracis by multiplex PCR analysis of sequences on plasmids pXO1 and pXO2 and chromosomal DNA.通过对质粒pXO1和pXO2以及染色体DNA上的序列进行多重PCR分析来鉴定和表征炭疽芽孢杆菌。
FEMS Microbiol Lett. 1996 Nov 15;145(1):9-16. doi: 10.1111/j.1574-6968.1996.tb08548.x.
5
[Application of the multiplex PCR and PCR-RFLP method in the identification of the Bacillus anthracis].多重聚合酶链反应和聚合酶链反应-限制性片段长度多态性方法在炭疽芽孢杆菌鉴定中的应用
Med Dosw Mikrobiol. 2005;57(3):277-85.
6
Genomic Characterization and Copy Number Variation of Plasmids pXO1 and pXO2 in a Historical Collection of 412 Strains.412株历史菌株中质粒pXO1和pXO2的基因组特征及拷贝数变异
mSystems. 2018 Aug 14;3(4). doi: 10.1128/mSystems.00065-18. eCollection 2018 Jul-Aug.
7
Multiplex PCR for species-level identification of Bacillus anthracis and detection of pXO1, pXO2, and related plasmids.多重 PCR 用于炭疽芽孢杆菌的种水平鉴定和 pXO1、pXO2 及相关质粒的检测。
Health Secur. 2015 Mar-Apr;13(2):122-9. doi: 10.1089/hs.2014.0056. Epub 2015 Mar 20.
8
In silico and in vitro evaluation of PCR-based assays for the detection of Bacillus anthracis chromosomal signature sequences.基于 PCR 的炭疽杆菌染色体特征序列检测分析方法的计算机模拟和体外评估。
Virulence. 2013 Nov 15;4(8):671-85. doi: 10.4161/viru.26288. Epub 2013 Sep 9.
9
Development and validation of a real-time quantitative PCR assay for rapid identification of Bacillus anthracis in environmental samples.开发和验证一种实时定量 PCR 检测方法，用于快速鉴定环境样本中的炭疽芽孢杆菌。
Appl Microbiol Biotechnol. 2010 Nov;88(5):1179-92. doi: 10.1007/s00253-010-2848-0. Epub 2010 Sep 9.
10
Identification of Bacillus anthracis by multiprobe microarray hybridization.通过多探针微阵列杂交鉴定炭疽芽孢杆菌。
Diagn Microbiol Infect Dis. 2004 Jul;49(3):163-71. doi: 10.1016/j.diagmicrobio.2004.03.015.

引用本文的文献

1
Multi-Functional Silver Nanoparticles for High-Throughput Endospore Sensing.多功能银纳米颗粒用于高通量内孢子感应。
Biosensors (Basel). 2022 Jan 25;12(2):68. doi: 10.3390/bios12020068.

本文引用的文献

1
Detection of Spp. and in Biological Samples by SYBR Green I and TaqMan Probe-based Real-time PCRs.通过SYBR Green I和基于TaqMan探针的实时聚合酶链反应检测生物样品中的特定物种和（此处原文不完整，缺少具体内容）
J Vet Res. 2017 Dec 27;61(4):427-432. doi: 10.1515/jvetres-2017-0069. eCollection 2017 Dec.
2
Pathogenicity, population genetics and dissemination of Bacillus anthracis.炭疽杆菌的致病性、种群遗传学和传播。
Infect Genet Evol. 2018 Oct;64:115-125. doi: 10.1016/j.meegid.2018.06.024. Epub 2018 Jun 20.
3
Sensitive and Specific Recombinase Polymerase Amplification Assays for Fast Screening, Detection, and Identification of Bacillus anthracis in a Field Setting.
用于在野外环境中快速筛选、检测和鉴定炭疽芽孢杆菌的灵敏且特异的重组酶聚合酶扩增检测方法。
Appl Environ Microbiol. 2018 May 17;84(11). doi: 10.1128/AEM.00506-18. Print 2018 Jun 1.
4
Historical evolution of human anthrax from occupational disease to potentially global threat as bioweapon.人类炭疽病的历史演变：从职业病到生物武器的潜在全球威胁
Environ Int. 2015 Dec;85:133-46. doi: 10.1016/j.envint.2015.09.009. Epub 2015 Sep 19.
5
Anthrax lethal and edema toxins in anthrax pathogenesis.炭疽致病过程中的炭疽致死毒素和水肿毒素。
Trends Microbiol. 2014 Jun;22(6):317-25. doi: 10.1016/j.tim.2014.02.012. Epub 2014 Mar 27.
6
Anthrax in animals.动物炭疽病。
Mol Aspects Med. 2009 Dec;30(6):481-9. doi: 10.1016/j.mam.2009.08.004. Epub 2009 Aug 31.
7
Bacillus anthracis physiology and genetics.炭疽芽孢杆菌生理学与遗传学。
Mol Aspects Med. 2009 Dec;30(6):386-96. doi: 10.1016/j.mam.2009.07.004. Epub 2009 Aug 3.
8
The surface of Bacillus anthracis.炭疽杆菌的表面。
Mol Aspects Med. 2009 Dec;30(6):374-85. doi: 10.1016/j.mam.2009.07.001. Epub 2009 Jul 14.
9
Dr. Jekyll and Mr. Hyde: a short history of anthrax.《化身博士：炭疽简史》
Mol Aspects Med. 2009 Dec;30(6):347-55. doi: 10.1016/j.mam.2009.06.004. Epub 2009 Jul 3.
10
Real-time PCR assay for a unique chromosomal sequence of Bacillus anthracis.用于炭疽芽孢杆菌独特染色体序列的实时聚合酶链反应检测
J Clin Microbiol. 2004 Dec;42(12):5825-31. doi: 10.1128/JCM.42.12.5825-5831.2004.