Design multiplex PCR for detection of rapid and correct the metallobetalactamase

被引:2
作者
Khosravi, Mohamad Ali [1 ,2 ,3 ]
Najafi, Ali [4 ]
Esmaeili, Davoud [1 ,2 ,3 ]
机构
[1] Baqiyatallah Univ Med Sci, Dept Microbiol, Tehran, Iran
[2] Baqiyatallah Univ Med Sci, Appl Microbiol Res Ctr, Syst Biol & Poisonings Inst, Tehran, Iran
[3] Baqiyatallah Univ Med Sci, Appl Virol ResearchCtr, Tehran, Iran
[4] Baqiyatallah Univ Med Sci, Mol Biol Res Ctr, Syst Biol & Poisonings Inst, Tehran, Iran
来源
GENE REPORTS | 2019年 / 17卷
关键词
Multiplex PCR; Detection; Rapid; Correct; Metallaobeta lactamase; PSEUDOMONAS-AERUGINOSA; BETA-LACTAMASES; KLEBSIELLA-PNEUMONIAE; RESISTANCE;
D O I
10.1016/j.genrep.2019.100498
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
One of the most important health and medical problems in many parts of the world is the spread of infectious diseases and the emergence of extent drug-resistant and innovation such as ESBL. Identification of them requires newer methods. One of the methods for identifying antibiotic resistance classes is Multiplex PCR. The purpose of this study is to identify ESBLs resistant gene using Multiplex PCR technique. In this study, three pair primers for ESBL (TEM, AmpC, KPC) were designed with bioinformatics software. Then, with control positive gene samples set-up of PCR was performed. Finally, additional experiments were performed with negative control samples and 50 isolates of the Escherichia coli isolated from Baqiyatallah hospital. The specificity and sensitivity of designed primers were also investigated. The results of the study showed that primers designed for MBL (SIM, GIM, IND) genes were able to simultaneously identify positive samples containing these genes. The sensitivity of Multiplex PCR for ESBL genes was 1 x 10-14. Also, the results showed that genotypic and phenotypic assays of clinical isolates of Escherichia coli were compatible with each other and the designed primers had good sensitivity (1 pg) and specificity (100%). This study showed that a Multiplex PCR designed with a sensitivity of 1 x 10-14 and a 100% specificity can correctly and fatly detect ESBL genes. Therefore, can suggest appropriate drug and antibiotic resistance to physicians determine the correct drug choice.
引用
收藏
页数:7
相关论文
共 41 条
  • [1] Arjmand-Asl M., 2016, KAUMS J FEYZ, V20, P376
  • [2] Badri A.M., 2017, MOL BIOL, V7, P10
  • [3] Bahador A, 2013, J PURE APPL MICROBIO, V7, P1559
  • [4] AmpC disk test for detection of plasmid-mediated AmpC β-lactamases in Enterobactetiaceae lacking chromosomal AmpC β-lactamases
    Black, JA
    Moland, ES
    Thomson, KS
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (07) : 3110 - 3113
  • [5] Molecular mechanisms of antibiotic resistance
    Blair, Jessica M. A.
    Webber, Mark A.
    Baylay, Alison J.
    Ogbolu, David O.
    Piddock, Laura J. V.
    [J]. NATURE REVIEWS MICROBIOLOGY, 2015, 13 (01) : 42 - 51
  • [6] Extended-spectrum β-lactamases in the 21st century:: Characterization, epidemiology, and detection of this important resistance threat
    Bradford, PA
    [J]. CLINICAL MICROBIOLOGY REVIEWS, 2001, 14 (04) : 933 - 951
  • [7] Bush K., 2016, COLD SPRING HARB PER, V21
  • [8] Bench-to-bedside review: The role of β-lactamases in antibiotic-resistant Gram-negative infections
    Bush, Karen
    [J]. CRITICAL CARE, 2010, 14 (03)
  • [9] Diversity of SHV and TEM β-lactamases in Klebsiella pneumoniae:: Gene evolution in northern Taiwan and two novel β-lactamases, SHV-25 and SHV-26
    Chang, FY
    Siu, LK
    Fung, CP
    Huang, MH
    Ho, M
    [J]. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 2001, 45 (09) : 2407 - 2413
  • [10] Recent Advances in Understanding Enteric Pathogenic Escherichia coli
    Croxen, Matthew A.
    Law, Robyn J.
    Scholz, Roland
    Keeney, Kristie M.
    Wlodarska, Marta
    Finlay, B. Brett
    [J]. CLINICAL MICROBIOLOGY REVIEWS, 2013, 26 (04) : 822 - 880