Accurate detection of methicillin-resistant staphylococcus aureus in mixtures by use of single-bacterium duplex droplet digital pcr
文献类型：期刊论文
| 作者 | Luo, Jun1,2; Li, Junhua1; Yang, Hang1; Yu, Junping1; Wei, Hongping1 |
| 刊名 | Journal of clinical microbiology
 |
| 出版日期 | 2017-10-01 |
| 卷号 | 55期号:10页码:2946-2955 |
| ISSN号 | 0095-1137 |
| 关键词 | Methicillin-resistant staphylococcus aureus Single-bacterium duplex droplet digital pcr Temperature-controlled lysis Chimeric lysin clyh Mrsa index ratios |
| DOI | 10.1128/jcm.00716-17 |
| 通讯作者 | Wei, hongping(hpwei@wh.iov.cn) |
| 英文摘要 | Accurate and rapid identification of methicillin-resistant staphylococcus aureus (mrsa) is needed to screen mrsa carriers and improve treatment. the current widely used duplex pcr methods are not able to differentiate mrsa from coexisting methicillin-susceptible s. aureus (mssa) or other methicillin-resistant staphylococci. in this study, we aimed to develop a direct method for accurate and rapid detection of mrsa in clinical samples from open environments, such as nasal swabs. the new molecular assay is based on detecting the cooccurrence of nuc and meca markers in a single bacterial cell by utilizing droplet digital pcr (ddpcr) with the chimeric lysin clyh for cell lysis. the method consists of (i) dispersion of an intact single bacterium into nanoliter droplets, (ii) temperature-controlled release of genomic dna (gdna) by clyh at 37 degrees c, and (iii) amplification and detection of the markers (nuc and meca) using standard taqman chemistries with ddpcr. results were analyzed based on mrsa index ratios used for indicating the presence of the duplexpositive markers in droplets. the method was able to achieve an absolute limit of detection (lod) of 2,900 cfu/ml for mrsa in nasal swabs spiked with excess amounts of escherichia coli, mssa, and other meca-positive bacteria within 4 h. initial testing of 104 nasal swabs showed that the method had 100% agreement with the standard culture method, while the normal duplex qpcr method had only about 87.5% agreement. the single-bacterium duplex ddpcr assay is rapid and powerful for more accurate detection of mrsa directly from clinical specimens. |
| WOS关键词 | REAL-TIME PCR ; RAPID DETECTION ; RISK-FACTORS ; MRSA ; ASSAY ; INFECTIONS ; SPECIMENS ; REGIONS ; IDENTIFICATION ; PERFORMANCE |
| WOS研究方向 | Microbiology |
| WOS类目 | Microbiology |
| 语种 | 英语 |
| 出版者 | AMER SOC MICROBIOLOGY |
| WOS记录号 | WOS:000411617400010 |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2373390 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Wei, Hongping |
| 作者单位 | 1.Chinese Acad Sci, Wuhan Inst Virol, Key Lab Special Pathogens & Biosafety, Wuhan, Hubei, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China |
| 推荐引用方式 GB/T 7714 | Luo, Jun,Li, Junhua,Yang, Hang,et al. Accurate detection of methicillin-resistant staphylococcus aureus in mixtures by use of single-bacterium duplex droplet digital pcr[J]. Journal of clinical microbiology,2017,55(10):2946-2955. |
| APA | Luo, Jun,Li, Junhua,Yang, Hang,Yu, Junping,&Wei, Hongping.(2017).Accurate detection of methicillin-resistant staphylococcus aureus in mixtures by use of single-bacterium duplex droplet digital pcr.Journal of clinical microbiology,55(10),2946-2955. |
| MLA | Luo, Jun,et al."Accurate detection of methicillin-resistant staphylococcus aureus in mixtures by use of single-bacterium duplex droplet digital pcr".Journal of clinical microbiology 55.10(2017):2946-2955. |
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来源：武汉病毒研究所
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