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| Development of a Rapid TaqMan Real-time PCR Assay for Detection of NDM-1 |
| DU Xin-ying, WANG Zhou-jia, WANG Yu-fei, QIU Shao-fu, YUAN Jing, SONG Hong-bin, SUN Yan-song, CHEN Ze-liang, HUANG Liu-yu |
| Institute of Disease Control and Prevention, Academy of Military Medical Science, Beijing 100071, China |
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Abstract The gene for NDM-1 (New Delhi metallo-beta-lactamase 1) is one member of a large gene family that encodes beta-lactamase enzymes. Bacteria that produce NDM-1 often referred to in the news media as "superbugs" because these bacterial are usually resistant to most antibiotics and infections caused by them are difficult to treat. NDM-1 gene was used for design of PCR primers and Taqman probe. Then a real-time quantitative PCR array for rapid detection of bacteria that produce NDM-1 was developed and its specifcity, sensitvity and repeatability were tested. Simulated urine specimens were used to assess the assay. A linear relationship was consistently obtained for input loads of 5×100~5×108 copies per assay. And the sensitivity of constructed assay method was 5 copies per reaction. No products were observed for strains of Enterobacteriaceae and only positive controls showed positive amplifications, indicating that the PCR assay is specific. The detection limit for simulated urine specimens was 10 CFU. These results indicated that the TaqMan real-time PCR assay described here is specific, sensitive and rapid for the detection of bacteria that produce NDM-1, and this assay could be used to detect the clinical specimens.
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Received: 11 January 2011
Published: 28 June 2011
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Cite this article:
DU Xin-ying, WANG Zhou-jia, WANG Yu-fei, QIU Shao-fu, YUAN Jing, SONG Hong-bin, SUN Yan-song, CHEN Ze-liang, HUANG Liu-yu. Development of a Rapid TaqMan Real-time PCR Assay for Detection of NDM-1. China Biotechnology, 2011, 31(06): 81-85.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I06/81
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| [1] Kamarasamy K, Toleman M A, Walsh T R, et al. Emergence of a new antibiotic resistance in India, Pakistan, and the UK: a prospective survey. Lancet Infect Dis, 2010, 10(9): 597-602.
[2] Struelens M J, Monnet D L, Magiorakos A P, et al. New Delhi metallo-beta-lactamase 1-producing Enterobacteriaceae: emergence and response in Europe. Euro Surveill, 2010, 15(46): 19716.
[3] Arya S C, Agarwal N. International travel with acquisition of multi-drug resistant Gram negative bacteria containing the New Delhi metallo-beta-lactamase gene, bla(NDM-1). Travel Med Infect Dis, 2011, 9(1): 47-48.
[4] Mulvey M R, Grant J M, Plewes K, et al. New Delhi metallo-β-lactamase in Klebsiella pneumoniae and Escherichia coli. Canada Emerg Infect Dis, 2011, 17(1): 103-106.
[5] Marra A. NDM-1: a local clone emerges with worldwide aspirations. Future Microbiol, 2011, 6: 137-141.
[6] Moellering R C Jr. NDM-1—a cause for worldwide concern. N Engl J Med, 2010, 363(25): 2377-2379.
[7] 杜昕颖, 孙宏迪, 王玉飞, 等. O1群霍乱弧菌实时荧光定量PCR快速检测方法的建立. 生物技术通讯, 2010, 21(5): 686-690. Du X Y, Sun H D, Wang Y F, et al. Letters in Biotechnology, 2010, 21(5): 686-690.
[8] Yong D, Toleman M A, Giske C G, et al. Characterization of a new metallo-beta-lactamase gene, bla (NDM-1), and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob Agents Chemother, 2009, 53: 5046-5054.
[9] Pillai D R, McGeer A, Low D E. New Delhi metallo-β-lactamase-1 in Enterobacteriaceae: emerging resistance. CMAJ, 2011, 183(1): 59-64.
[10] Espy M J, Uhl J R, Sloan L M, et al. Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clin Microbiol Rev, 2006, 19: 165-256. |
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