对预混试剂进行脱氧核糖核酸酶预处理可提高通用16S核糖体RNA基因聚合酶链反应结果的有效性。
DNase pretreatment of master mix reagents improves the validity of universal 16S rRNA gene PCR results.
作者信息
Heininger Alexandra, Binder Marlies, Ellinger Andreas, Botzenhart Konrad, Unertl Klaus, Döring Gerd
机构信息
Klinik für Anästhesiologie und Transfusionsmedizin, Abteilung für Anästhesiologie, Eberhard-Karls-Universität Tübingen, D-72074 Tübingen, Germany.
出版信息
J Clin Microbiol. 2003 Apr;41(4):1763-5. doi: 10.1128/JCM.41.4.1763-1765.2003.
Abstract
DNase I pretreatment of 16S rRNA gene PCR reagents was tested. The DNase I requirement for the elimination of false-positive results varied between 0.1 and 70 IU per master mix depending on the applied Taq polymerase. PCR sensitivity was mostly maintained when 0.1 IU of DNase I was used.
摘要
对16S rRNA基因PCR试剂进行了DNase I预处理测试。根据所使用的Taq聚合酶不同，消除假阳性结果所需的DNase I量在每个反应预混液0.1至70 IU之间变化。当使用0.1 IU的DNase I时，PCR灵敏度大多能得以维持。
相似文献
1
DNase pretreatment of master mix reagents improves the validity of universal 16S rRNA gene PCR results.对预混试剂进行脱氧核糖核酸酶预处理可提高通用16S核糖体RNA基因聚合酶链反应结果的有效性。
J Clin Microbiol. 2003 Apr;41(4):1763-5. doi: 10.1128/JCM.41.4.1763-1765.2003.
2
Reagent decontamination to eliminate false-positives in Escherichia coli qPCR.用于消除大肠杆菌定量聚合酶链反应中假阳性的试剂去污处理
J Microbiol Methods. 2008 Mar;72(3):275-82. doi: 10.1016/j.mimet.2007.12.011. Epub 2007 Dec 31.
3
Optimization of real-time PCR assay for rapid and sensitive detection of eubacterial 16S ribosomal DNA in platelet concentrates.优化实时聚合酶链反应检测方法以快速灵敏地检测浓缩血小板中的真细菌16S核糖体DNA
J Clin Microbiol. 2003 Oct;41(10):4796-8. doi: 10.1128/JCM.41.10.4796-4798.2003.
4
Activity and DNA contamination of commercial polymerase chain reaction reagents for the universal 16S rDNA real-time polymerase chain reaction detection of bacterial pathogens in blood.商用聚合酶链反应试剂对血液中细菌病原体的通用 16S rDNA 实时聚合酶链反应检测的活性和 DNA 污染。
Diagn Microbiol Infect Dis. 2010 Jan;66(1):41-9. doi: 10.1016/j.diagmicrobio.2008.07.011. Epub 2008 Aug 21.
5
Contamination and sensitivity issues with a real-time universal 16S rRNA PCR.实时通用16S rRNA聚合酶链反应的污染和敏感性问题。
J Clin Microbiol. 2000 May;38(5):1747-52. doi: 10.1128/JCM.38.5.1747-1752.2000.
6
Effect of comprehensive validation of the template isolation procedure on the reliability of bacteraemia detection by a 16S rRNA gene PCR.模板分离程序的全面验证对通过16S rRNA基因PCR检测菌血症可靠性的影响。
Clin Microbiol Infect. 2004 May;10(5):452-8. doi: 10.1111/j.1469-0691.2004.00877.x.
7
DNase I treatment of Taq DNA polymerase for complete PCR decontamination.用脱氧核糖核酸酶I处理Taq DNA聚合酶以实现完全的聚合酶链式反应（PCR）污染清除。
Biotechniques. 1992 Oct;13(4):520.
8
Decontamination of polymerase chain reaction reagents for detection of low concentrations of 16S rRNA genes.用于检测低浓度16S rRNA基因的聚合酶链反应试剂的净化处理
Mol Biotechnol. 1997 Jun;7(3):207-16. doi: 10.1007/BF02740812.
9
Nonspecific PCR amplification of the 16S rRNA gene segment in different bacteria by use of primers specific for Campylobacter, Arcobacter, and Helicobacter spp.使用针对弯曲杆菌属、弓形杆菌属和幽门螺杆菌属的特异性引物对不同细菌中的16S rRNA基因片段进行非特异性PCR扩增。
J Clin Microbiol. 2007 Apr;45(4):1376-7. doi: 10.1128/JCM.02559-06.
10
Comparison of different decontamination methods for reagents to detect low concentrations of bacterial 16S DNA by real-time-PCR.通过实时聚合酶链反应检测低浓度细菌16S DNA的试剂的不同去污方法比较
Mol Biotechnol. 2002 Nov;22(3):231-42. doi: 10.1385/MB:22:3:231.
引用本文的文献
1
Validating digital polymerase chain reaction for 16S rRNA gene amplification from low biomass environmental samples.验证数字聚合酶链反应用于从低生物量环境样本中扩增16S rRNA基因。
ISME Commun. 2025 Jul 9;5(1):ycaf115. doi: 10.1093/ismeco/ycaf115. eCollection 2025 Jan.
2
A Clinico-Microbiological Study of Prosthetic Joint Infections in an Indian Tertiary Care Hospital: Role of Universal 16S rRNA Gene Polymerase Chain Reaction and Sequencing in Diagnosis.印度一家三级护理医院人工关节感染的临床微生物学研究：通用16S rRNA基因聚合酶链反应和测序在诊断中的作用
Indian J Orthop. 2019 Sep-Oct;53(5):646-654. doi: 10.4103/ortho.IJOrtho_551_18.
3
Utility of 16S rRNA PCR in the Synovial Fluid for the Diagnosis of Prosthetic Joint Infection.16S rRNA聚合酶链反应在滑液中诊断人工关节感染的效用
Ann Lab Med. 2018 Nov;38(6):610-612. doi: 10.3343/alm.2018.38.6.610.
4
A broad-range PCR technique for the diagnosis of infective endocarditis.一种用于诊断感染性心内膜炎的广谱聚合酶链反应技术。
Braz J Microbiol. 2018 Jul-Sep;49(3):534-543. doi: 10.1016/j.bjm.2017.03.019. Epub 2018 Feb 9.
5
Removal of Contaminant DNA by Combined UV-EMA Treatment Allows Low Copy Number Detection of Clinically Relevant Bacteria Using Pan-Bacterial Real-Time PCR.通过紫外线与乙二胺四乙酸联合处理去除污染DNA，可使用泛细菌实时PCR对临床相关细菌进行低拷贝数检测。
PLoS One. 2015 Jul 14;10(7):e0132954. doi: 10.1371/journal.pone.0132954. eCollection 2015.
6
Reagent and laboratory contamination can critically impact sequence-based microbiome analyses.试剂和实验室污染会严重影响基于序列的微生物组分析。
BMC Biol. 2014 Nov 12;12:87. doi: 10.1186/s12915-014-0087-z.
7
Expression of CD64 on neutrophils can be used to predict the severity of bloodstream infection before broad range 16S rRNA PCR.中性粒细胞上CD64的表达可用于在进行广泛的16S rRNA PCR之前预测血流感染的严重程度。
Folia Microbiol (Praha). 2015 Mar;60(2):111-8. doi: 10.1007/s12223-014-0346-y. Epub 2014 Sep 26.
8
Non-culture-based methods to diagnose bloodstream infection: Does it work?非基于培养的血流感染诊断方法：可行吗？
Eur J Microbiol Immunol (Bp). 2013 Jun;3(2):97-104. doi: 10.1556/EuJMI.3.2013.2.2. Epub 2013 Jun 5.
9
Use of staged molecular analysis to determine causes of unexplained central nervous system infections.采用阶段性分子分析确定不明原因中枢神经系统感染的病因。
Emerg Infect Dis. 2013;19(9):1470-7. doi: 10.3201/eid1909.130474.
10
Evaluation of an ethidium monoazide-enhanced 16S rDNA real-time polymerase chain reaction assay for bacterial screening of platelet concentrates and comparison with automated culture.评价一种基于吖啶橙增强的 16S rDNA 实时聚合酶链反应检测法，用于血小板浓缩物的细菌筛选，并与自动化培养进行比较。
Transfusion. 2014 Mar;54(3 Pt 2):870-8. doi: 10.1111/trf.12256. Epub 2013 May 23.
本文引用的文献
1
Detection of nucleic acid sequences from bacterial species with molecular genetic methods.用分子遗传学方法检测细菌物种的核酸序列。
Transfus Sci. 2000 Aug;23(1):21-7. doi: 10.1016/s0955-3886(00)00051-5.
2
DNase I activity retained after heat inactivation in standard buffer.在标准缓冲液中热灭活后保留的脱氧核糖核酸酶I活性。
Biotechniques. 2000 Jul;29(1):38-40, 42. doi: 10.2144/00291bm05.
3
Use of PCR with universal primers and restriction endonuclease digestions for detection and identification of common bacterial pathogens in cerebrospinal fluid.使用通用引物的聚合酶链反应（PCR）和限制性内切酶消化法检测和鉴定脑脊液中的常见细菌病原体。
J Clin Microbiol. 2000 Jun;38(6):2076-80. doi: 10.1128/JCM.38.6.2076-2080.2000.
4
Contamination and sensitivity issues with a real-time universal 16S rRNA PCR.实时通用16S rRNA聚合酶链反应的污染和敏感性问题。
J Clin Microbiol. 2000 May;38(5):1747-52. doi: 10.1128/JCM.38.5.1747-1752.2000.
5
Rapid diagnosis of bacteremia by universal amplification of 23S ribosomal DNA followed by hybridization to an oligonucleotide array.通过对23S核糖体DNA进行通用扩增，随后与寡核苷酸阵列杂交来快速诊断菌血症。
J Clin Microbiol. 2000 Feb;38(2):781-8. doi: 10.1128/JCM.38.2.781-788.2000.
6
Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion.通过限制性内切酶消化从Taq DNA聚合酶中去除细菌DNA。
J Clin Microbiol. 1999 Oct;37(10):3402-4. doi: 10.1128/JCM.37.10.3402-3404.1999.
7
PCR and blood culture for detection of Escherichia coli bacteremia in rats.用于检测大鼠大肠杆菌菌血症的聚合酶链反应（PCR）和血培养
J Clin Microbiol. 1999 Aug;37(8):2479-82. doi: 10.1128/JCM.37.8.2479-2482.1999.
8
The use of polymerase chain reaction to detect septicemia in critically ill patients.使用聚合酶链反应检测重症患者的败血症。
Crit Care Med. 1999 May;27(5):937-40. doi: 10.1097/00003246-199905000-00029.
9
Detection of bacteraemia in patients with fever and neutropenia using 16S rRNA gene amplification by polymerase chain reaction.采用聚合酶链反应通过16S rRNA基因扩增检测发热伴中性粒细胞减少患者的菌血症
Eur J Clin Microbiol Infect Dis. 1998 Apr;17(4):247-53. doi: 10.1007/BF01699981.
10
Molecular diagnosis of bacterial endocarditis by broad-range PCR amplification and direct sequencing.通过广谱PCR扩增和直接测序对细菌性心内膜炎进行分子诊断。
J Clin Microbiol. 1997 Nov;35(11):2733-9. doi: 10.1128/jcm.35.11.2733-2739.1997.
Zotero 插件