دورية أكاديمية
Express Yourself: Quantitative Real-Time PCR Assays for Rapid Chromosomal Antimicrobial Resistance Detection in Pseudomonas aeruginosa.
العنوان: | Express Yourself: Quantitative Real-Time PCR Assays for Rapid Chromosomal Antimicrobial Resistance Detection in Pseudomonas aeruginosa. |
---|---|
المؤلفون: | Madden DE; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Olagoke O; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Baird T; Sunshine Coast Health Institute, Birtinya, Queensland, Australia.; Respiratory Department, Sunshine Coast University Hospital, Birtinya, Queensland, Australia., Neill J; Sunshine Coast Health Institute, Birtinya, Queensland, Australia.; Respiratory Department, Sunshine Coast University Hospital, Birtinya, Queensland, Australia., Ramsay KA; Child Health Research Centre, The University of Queenslandgrid.1003.2, South Brisbane, Queensland, Australia., Fraser TA; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Bell SC; Child Health Research Centre, The University of Queenslandgrid.1003.2, South Brisbane, Queensland, Australia.; Adult Cystic Fibrosis Centre, The Prince Charles Hospital, Chermside, Queensland, Australia.; Translational Research Institute, Woolloongabba, Queensland, Australia., Sarovich DS; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Price EP; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia. |
المصدر: | Antimicrobial agents and chemotherapy [Antimicrob Agents Chemother] 2022 May 17; Vol. 66 (5), pp. e0020422. Date of Electronic Publication: 2022 Apr 25. |
نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 0315061 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-6596 (Electronic) Linking ISSN: 00664804 NLM ISO Abbreviation: Antimicrob Agents Chemother Subsets: MEDLINE |
أسماء مطبوعة: | Original Publication: Washington, American Society for Microbiology |
مواضيع طبية MeSH: | Cystic Fibrosis*/complications , Pseudomonas Infections*/microbiology, Anti-Bacterial Agents/therapeutic use ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/metabolism ; Carbapenems/therapeutic use ; Drug Resistance, Bacterial ; Humans ; Membrane Transport Proteins/genetics ; Microbial Sensitivity Tests ; Pseudomonas aeruginosa ; Real-Time Polymerase Chain Reaction |
مستخلص: | The rise of antimicrobial-resistant (AMR) bacteria is a global health emergency. One critical facet of tackling this epidemic is more rapid AMR diagnosis in serious multidrug-resistant pathogens like Pseudomonas aeruginosa. Here, we designed and then validated two multiplex quantitative real-time PCR (qPCR) assays to simultaneously detect differential expression of the resistance-nodulation-division efflux pumps MexAB-OprM, MexCD-OprJ, MexEF-OprN, and MexXY-OprM, the AmpC β-lactamase, and the porin OprD, which are commonly associated with chromosomally encoded AMR. Next, qPCRs were tested on 15 sputa from 11 participants with P. aeruginosa respiratory infections to determine AMR profiles in vivo . We confirmed multiplex qPCR testing feasibility directly on sputa, representing a key advancement in in vivo AMR diagnosis. Notably, comparison of sputa with their derived isolates grown in Luria-Bertani broth (±2.5% NaCl) or a 5-antibiotic cocktail showed marked expression differences, illustrating the difficulty in replicating in vivo expression profiles in vitro . Cystic fibrosis sputa showed significantly reduced mexE and mexY expression compared with chronic obstructive pulmonary disease sputa, despite harboring fluoroquinolone- and aminoglycoside-resistant strains, indicating that these loci do not contribute to AMR in vivo. oprD was also significantly downregulated in cystic fibrosis sputa, even in the absence of contemporaneous carbapenem use, suggesting a common adaptive trait in chronic infections that may affect carbapenem efficacy. Sputum ampC expression was highest in participants receiving carbapenems (6.7 to 15×), some of whom were simultaneously receiving cephalosporins, the latter of which would be rendered ineffective by the upregulated ampC . Our qPCR assays provide valuable insights into the P. aeruginosa resistome, and their use on clinical specimens will permit timely treatment alterations that will improve patient outcomes and antimicrobial stewardship measures. |
References: | Mol Microbiol. 1996 Aug;21(4):713-24. (PMID: 8878035) Proc Natl Acad Sci U S A. 2011 Oct 11;108(41):17165-70. (PMID: 21969582) Microb Genom. 2022 Feb;8(2):. (PMID: 35113778) EBioMedicine. 2021 Jan;63:103152. (PMID: 33285499) Mol Microbiol. 2002 Mar;43(6):1641-50. (PMID: 11952911) Antimicrob Agents Chemother. 1998 Aug;42(8):1938-43. (PMID: 9687387) Infect Genet Evol. 2014 Jun;24:187-92. (PMID: 24694825) Antimicrob Agents Chemother. 2008 Dec;52(12):4478-82. (PMID: 18838593) Diagn Microbiol Infect Dis. 2010 Sep;68(1):20-7. (PMID: 20727465) J Antimicrob Chemother. 2017 Sep 1;72(9):2489-2495. (PMID: 28535274) Future Microbiol. 2018 Sep;13:1403-1418. (PMID: 30256166) Res Microbiol. 1992 Oct;143(8):785-90. (PMID: 1298031) J Clin Microbiol. 2019 Feb 27;57(3):. (PMID: 30381421) J Bacteriol. 1999 Oct;181(20):6300-5. (PMID: 10515918) Trends Microbiol. 2011 Aug;19(8):419-26. (PMID: 21664819) Lancet Infect Dis. 2013 Dec;13(12):1057-98. (PMID: 24252483) J Am Chem Soc. 2015 Jan 14;137(1):190-200. (PMID: 25495032) Diagn Microbiol Infect Dis. 2009 Feb;63(2):127-31. (PMID: 19026507) mBio. 2015 Nov 24;6(6):e01796-15. (PMID: 26604259) Int J Antimicrob Agents. 2019 May;53(5):620-628. (PMID: 30664925) Clin Microbiol Infect. 2018 Apr;24(4):355-360. (PMID: 29117578) Int J Antimicrob Agents. 2003 May;21(5):409-13. (PMID: 12727072) Nat Commun. 2021 Apr 28;12(1):2460. (PMID: 33911082) Antimicrob Agents Chemother. 2000 Dec;44(12):3322-7. (PMID: 11083635) Int J Antimicrob Agents. 2012 Aug;40(2):168-72. (PMID: 22633564) Antimicrob Agents Chemother. 2001 Feb;45(2):480-4. (PMID: 11158744) J Clin Microbiol. 2005 Oct;43(10):5085-90. (PMID: 16207967) Int J Antimicrob Agents. 2015 May;45(5):529-32. (PMID: 25735764) Front Microbiol. 2011 Apr 05;2:65. (PMID: 21747788) Microb Genom. 2019 Oct;5(10):. (PMID: 31617838) Philos Trans R Soc Lond B Biol Sci. 2015 Jun 5;370(1670):20140082. (PMID: 25918440) Antimicrob Agents Chemother. 1996 Sep;40(9):2021-8. (PMID: 8878574) Drugs Context. 2018 May 29;7:212527. (PMID: 29872449) Clin Infect Dis. 2008 Dec 15;47(12):1534-6. (PMID: 19025364) Antimicrob Agents Chemother. 2020 Apr 21;64(5):. (PMID: 32071060) Microbiology (Reading). 2013 Oct;159(Pt 10):2058-2073. (PMID: 23924707) Antimicrob Resist Infect Control. 2019 Aug 13;8:137. (PMID: 31417673) Microbiology (Reading). 2002 Jan;148(Pt 1):257-266. (PMID: 11782518) Antimicrob Agents Chemother. 2021 May 18;65(6):. (PMID: 33781999) Antimicrob Agents Chemother. 2018 Jan 25;62(2):. (PMID: 29133559) Antimicrob Agents Chemother. 2015 Jan;59(1):427-36. (PMID: 25367914) PLoS One. 2012;7(5):e37723. (PMID: 22624061) Bioinformatics. 2018 Mar 1;34(5):867-868. (PMID: 29096012) Eur Respir Rev. 2021 Sep 15;30(161):. (PMID: 34526313) Mol Microbiol. 2016 Jun;100(5):749-58. (PMID: 26844397) Infect Drug Resist. 2019 Feb 08;12:261-272. (PMID: 30804676) Drug Resist Updat. 2019 May;44:100640. (PMID: 31492517) Trends Microbiol. 2016 May;24(5):327-337. (PMID: 26946977) Future Microbiol. 2016;11(1):81-92. (PMID: 26674470) Antimicrob Agents Chemother. 2016 Nov 21;60(12):7415-7423. (PMID: 27736752) J Med Microbiol. 2022 Oct;71(10):. (PMID: 36301593) J Med Microbiol. 2010 Sep;59(Pt 9):1089-1100. (PMID: 20522626) J Antimicrob Chemother. 2020 Dec 1;75(12):3491-3500. (PMID: 32780112) J Vis Exp. 2012 Jun 05;(64):e3857. (PMID: 22711026) Int J Antimicrob Agents. 2016 Dec;48(6):740-743. (PMID: 28128097) Front Microbiol. 2019 Apr 01;10:539. (PMID: 30988669) Mol Microbiol. 1997 Jan;23(2):345-54. (PMID: 9044268) Antimicrob Agents Chemother. 2018 Oct 24;62(11):. (PMID: 30201819) Antimicrob Agents Chemother. 2009 Jun;53(6):2522-31. (PMID: 19332674) Antimicrob Agents Chemother. 2006 May;50(5):1633-41. (PMID: 16641429) Antimicrob Agents Chemother. 2005 Nov;49(11):4733-8. (PMID: 16251318) mBio. 2021 Aug 31;12(4):e0047321. (PMID: 34281392) Antimicrob Agents Chemother. 2011 Feb;55(2):508-14. (PMID: 21078928) Clin Microbiol Rev. 1995 Oct;8(4):557-84. (PMID: 8665470) Methods. 2001 Dec;25(4):402-8. (PMID: 11846609) BMC Microbiol. 2010 Dec 30;10:328. (PMID: 21192796) Antimicrob Agents Chemother. 2010 May;54(5):2219-24. (PMID: 20194693) Dev Cell. 2004 Nov;7(5):745-54. (PMID: 15525535) Clin Microbiol Infect. 2004 Oct;10(10):877-83. (PMID: 15373880) Appl Environ Microbiol. 2004 Oct;70(10):6188-96. (PMID: 15466566) Front Microbiol. 2012 Nov 28;3:408. (PMID: 23233851) J Clin Microbiol. 2005 Dec;43(12):5945-9. (PMID: 16333080) Antimicrob Agents Chemother. 2006 May;50(5):1780-7. (PMID: 16641450) PLoS One. 2017 Mar 8;12(3):e0172179. (PMID: 28273168) Antimicrob Agents Chemother. 2021 Jun 17;65(7):e0269620. (PMID: 33875431) Int J Antimicrob Agents. 2005 Apr;25(4):290-5. (PMID: 15784307) Clin Microbiol Rev. 2019 Aug 28;32(4):. (PMID: 31462403) J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Aug 1;877(23):2224-34. (PMID: 18929516) Arch Med Res. 2005 Nov-Dec;36(6):697-705. (PMID: 16216651) Genome Biol. 2019 Nov 28;20(1):257. (PMID: 31779668) Antimicrob Agents Chemother. 2011 May;55(5):1906-11. (PMID: 21357294) PLoS One. 2013 Apr 29;8(4):e62774. (PMID: 23658651) PLoS Pathog. 2020 Sep 30;16(9):e1008357. (PMID: 32997707) Antimicrob Agents Chemother. 2009 May;53(5):1987-97. (PMID: 19258280) J Antimicrob Chemother. 2010 Jul;65(7):1540-2. (PMID: 20435778) Antimicrob Agents Chemother. 2017 Oct 24;61(11):. (PMID: 28874376) J Cyst Fibros. 2022 Jan;21(1):e35-e43. (PMID: 33775602) Microb Drug Resist. 2003 Winter;9(4):323-8. (PMID: 15000738) |
فهرسة مساهمة: | Keywords: AMR; Pseudomonas aeruginosa; ampC; antibiotic; antibiotic resistance; assay development; diagnostics; efflux pumps; gene expression; genomics; in vivo expression technology; qPCR |
المشرفين على المادة: | 0 (Anti-Bacterial Agents) 0 (Bacterial Outer Membrane Proteins) 0 (Bacterial Proteins) 0 (Carbapenems) 0 (Membrane Transport Proteins) |
تواريخ الأحداث: | Date Created: 20220425 Date Completed: 20220519 Latest Revision: 20240828 |
رمز التحديث: | 20240828 |
مُعرف محوري في PubMed: | PMC9112894 |
DOI: | 10.1128/aac.00204-22 |
PMID: | 35467369 |
قاعدة البيانات: | MEDLINE |
كن أول من يترك تعليقا!