用于检测耐甲氧西林金黄色葡萄球菌中mecA基因的酶标寡核苷酸探针的研制
Development of enzyme-labeled oligonucleotide probe for detection of mecA gene in methicillin-resistant Staphylococcus aureus.
作者信息
Shimaoka M, Yoh M, Segawa A, Takarada Y, Yamamoto K, Honda T
机构信息
Research Institute for Microbial Diseases, Osaka University, Japan.
出版信息
J Clin Microbiol. 1994 Aug;32(8):1866-9. doi: 10.1128/jcm.32.8.1866-1869.1994.
Abstract
A DNA hybridization method with an enzyme-labeled oligonucleotide probe (mecA-ELONP) was developed to detect the methicillin-resistant gene (mecA) in methicillin-resistant Staphylococcus aureus. For rapid identification, bacterial colonies were transferred from agar plates directly onto nylon membranes. Lysis of cells, denaturation of DNA, and hybridization were performed on the membranes. These procedures required only 3 h for completion. The results obtained by this test closely corresponded with those obtained by determining the MICs of oxacillin against S. aureus. The results of the mecA-ELONP also correlated well with those of a commercially available PCR test. Thus, mecA-ELONP proved to be a reliable and convenient method for the rapid identification of methicillin-resistant S. aureus, which could be useful in clinical microbiology laboratories.
摘要
开发了一种用酶标记寡核苷酸探针（mecA-ELONP）的DNA杂交方法，用于检测耐甲氧西林金黄色葡萄球菌中的耐甲氧西林基因（mecA）。为了快速鉴定，将细菌菌落从琼脂平板直接转移到尼龙膜上。在膜上进行细胞裂解、DNA变性和杂交。这些步骤仅需3小时即可完成。该试验获得的结果与通过测定奥沙西林对金黄色葡萄球菌的最低抑菌浓度所获得的结果密切相符。mecA-ELONP的结果也与市售PCR试验的结果相关性良好。因此，mecA-ELONP被证明是一种可靠且方便的快速鉴定耐甲氧西林金黄色葡萄球菌的方法，这在临床微生物学实验室中可能会很有用。
相似文献
1
Development of enzyme-labeled oligonucleotide probe for detection of mecA gene in methicillin-resistant Staphylococcus aureus.用于检测耐甲氧西林金黄色葡萄球菌中mecA基因的酶标寡核苷酸探针的研制
J Clin Microbiol. 1994 Aug;32(8):1866-9. doi: 10.1128/jcm.32.8.1866-1869.1994.
2
Detection of the gene for toxic shock syndrome toxin 1 in Staphylococcus aureus by enzyme-labelled oligonucleotide probes.
J Med Microbiol. 1996 Mar;44(3):215-8. doi: 10.1099/00222615-44-3-215.
3
Detection of methicillin- and aminoglycoside-resistant genes and simultaneous identification of S. aureus using triplex real-time PCR Taqman assay.采用三重实时荧光定量PCR Taqman分析法检测耐甲氧西林和氨基糖苷类基因并同时鉴定金黄色葡萄球菌。
J Microbiol Methods. 2007 Jan;68(1):157-62. doi: 10.1016/j.mimet.2006.07.008. Epub 2006 Aug 28.
4
Comparison of phenotypic methods with mecA gene based polymerase chain reaction for Methicillin-resistant Staphylococcus aureus detection.基于mecA基因的聚合酶链反应与表型方法检测耐甲氧西林金黄色葡萄球菌的比较。
J Pak Med Assoc. 2020 Feb;70(2):276-280. doi: 10.5455/JPMA.290732.
5
Evaluation of a new 3-h hybridization method for detecting the mecA gene in Staphylococcus aureus and comparison with existing genotypic and phenotypic susceptibility testing methods.评估一种用于检测金黄色葡萄球菌中mecA基因的新型3小时杂交方法，并与现有的基因型和表型药敏试验方法进行比较。
J Antimicrob Chemother. 1999 Apr;43(4):467-75. doi: 10.1093/jac/43.4.467.
6
[Detection of methicillin resistance and identification of Staphylococcus spp. from positive blood culture bottles using the mecA and nucA genes with the LightCycler System].[使用LightCycler系统通过mecA和nucA基因检测耐甲氧西林情况并鉴定血培养阳性瓶中的葡萄球菌属]
Enferm Infecc Microbiol Clin. 2005 Apr;23(4):208-12. doi: 10.1157/13073146.
7
Unusual form of oxacillin resistance in methicillin-resistant Staphylococcus aureus clinical strains.耐甲氧西林金黄色葡萄球菌临床菌株中不寻常的苯唑西林耐药形式。
Diagn Microbiol Infect Dis. 2008 Aug;61(4):387-95. doi: 10.1016/j.diagmicrobio.2008.04.003. Epub 2008 May 27.
8
[Drug-resistance of staphylococcus aureus and detection of mecA gene in all strains isolated from children in Hangzhou].[杭州地区儿童分离株金黄色葡萄球菌耐药性及mecA基因检测]
Zhonghua Er Ke Za Zhi. 2006 May;44(5):360-3.
9
Testing of methicillin resistance by in vitro susceptibility and the presence of the mecA gene in clinical Staphylococcus aureus isolates in Finland.
Scand J Infect Dis. 1995;27(5):475-9. doi: 10.3109/00365549509047049.
10
Validation of multiplex PCR strategy for simultaneous detection and identification of methicillin resistant Staphylococcus aureus.用于同时检测和鉴定耐甲氧西林金黄色葡萄球菌的多重PCR策略的验证
Indian J Med Microbiol. 2008 Oct-Dec;26(4):361-4. doi: 10.4103/0255-0857.43580.
引用本文的文献
1
Rapid identification of staphylococcal strains from positive-testing blood culture bottles by internal transcribed spacer PCR followed by microchip gel electrophoresis.通过内转录间隔区聚合酶链反应继之以微芯片凝胶电泳从阳性血培养瓶中快速鉴定葡萄球菌菌株。
J Clin Microbiol. 2005 Mar;43(3):1149-57. doi: 10.1128/JCM.43.3.1149-1157.2005.
2
Molecular diagnostics of clinically important staphylococci.临床重要葡萄球菌的分子诊断
Folia Microbiol (Praha). 2004;49(4):353-86. doi: 10.1007/BF03354664.
3
Oxacillin susceptibility testing of staphylococci directly from Bactec Plus blood cultures by the BBL Crystal MRSA ID system.使用BBL Crystal MRSA ID系统对直接来自Bactec Plus血培养的葡萄球菌进行苯唑西林药敏试验。
J Clin Microbiol. 1999 Jun;37(6):2034-6. doi: 10.1128/JCM.37.6.2034-2036.1999.
本文引用的文献
1
Development of an enzyme-labeled oligonucleotide probe for the cholera toxin gene.霍乱毒素基因酶标寡核苷酸探针的研制
J Clin Microbiol. 1993 May;31(5):1312-4. doi: 10.1128/jcm.31.5.1312-1314.1993.
2
Concordant clonal delineation of methicillin-resistant Staphylococcus aureus by macrorestriction analysis and polymerase chain reaction genome fingerprinting.通过宏观限制性分析和聚合酶链反应基因组指纹图谱对耐甲氧西林金黄色葡萄球菌进行一致的克隆鉴定。
J Clin Microbiol. 1993 Aug;31(8):1964-70. doi: 10.1128/jcm.31.8.1964-1970.1993.
3
Occurrence of a beta-lactam-inducible penicillin-binding protein in methicillin-resistant staphylococci.耐甲氧西林葡萄球菌中β-内酰胺诱导型青霉素结合蛋白的出现。
Antimicrob Agents Chemother. 1985 May;27(5):851-7. doi: 10.1128/AAC.27.5.851.
4
Regulation of penicillin-binding protein activity: description of a methicillin-inducible penicillin-binding protein in Staphylococcus aureus.青霉素结合蛋白活性的调节：金黄色葡萄球菌中一种甲氧西林诱导型青霉素结合蛋白的描述。
Antimicrob Agents Chemother. 1985 May;27(5):828-31. doi: 10.1128/AAC.27.5.828.
5
Preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use as hybridization probes.寡脱氧核苷酸-碱性磷酸酶缀合物的制备及其作为杂交探针的用途。
Nucleic Acids Res. 1986 Aug 11;14(15):6115-28. doi: 10.1093/nar/14.15.6115.
6
Production of low-affinity penicillin-binding protein by low- and high-resistance groups of methicillin-resistant Staphylococcus aureus.耐甲氧西林金黄色葡萄球菌低抗性组和高抗性组产生低亲和力青霉素结合蛋白。
Antimicrob Agents Chemother. 1987 Sep;31(9):1307-11. doi: 10.1128/AAC.31.9.1307.
7
Expression of methicillin resistance in heterogeneous strains of Staphylococcus aureus.金黄色葡萄球菌异质菌株中耐甲氧西林的表达
Antimicrob Agents Chemother. 1986 Jan;29(1):85-92. doi: 10.1128/AAC.29.1.85.
8
Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion.通过基因融合在耐甲氧西林金黄色葡萄球菌中诱导型青霉素靶蛋白的进化
FEBS Lett. 1987 Aug 31;221(1):167-71. doi: 10.1016/0014-5793(87)80373-3.
9
Evaluation of a nonisotopically labeled oligonucleotide probe to detect the heat-stable enterotoxin gene of Escherichia coli by the DNA colony hybridization test.通过DNA菌落杂交试验评估一种非同位素标记的寡核苷酸探针以检测大肠杆菌热稳定肠毒素基因。
J Clin Microbiol. 1988 Apr;26(4):784-6. doi: 10.1128/jcm.26.4.784-786.1988.
10
The role of beta-lactamase in staphylococcal resistance to penicillinase-resistant penicillins and cephalosporins.β-内酰胺酶在葡萄球菌对耐青霉素酶青霉素和头孢菌素耐药性中的作用。
J Clin Microbiol. 1986 May;23(5):832-9. doi: 10.1128/jcm.23.5.832-839.1986.
Zotero 插件