Evaluation of a commercial multiplex real-time PCR panel for identification environmental water antibiotic-resistant bacteria in hemoculture bottles
Article Sidebar
Main Article Content
Abstract
Background: Antimicrobial resistance (AMR) is a major public health problem worldwide, and it is increasing, especially in developed and developing countries, including Thailand. Sepsis is a crisis and emergency causing the death of patients with a high mortality rate of 1 in 5 of the global population. Using inappropriate antimicrobial drugs to treat infections increases the chance of drug resistance in the microorganisms, and precise and rapid treatment is essential for patient management.
Objective: This cooperative study aims to evaluate a commercial multiplex real-time PCR panel, QIAstat-Dx BCID GN Plus AMR Panel, to detect antimicrobial-resistant genes in the bloodstream.
Materials and methods: Thirty bacterial samples were used to simulate bloodstream infections. The comparison between the QIAstat-Dx BCID GN Plus AMR Panel and the conventional PCR method was revealed.
Results: Both methods gave concordant results for 12 samples, while results for 15 were discordant. Three samples could not be detected using the QIAstat-Dx BCID GN Plus AMR Panel because ebc gene is not included. The QIAstat-Dx BCID GN Plus AMR Panel revealed a sensitivity, specificity, positive predictive value, and negative predictive value of 91.50%, 93.30%, 74.10%, and 98.50%, respectively.
Conclusion: The study indicated that the QIAstat-Dx BCID GN Plus AMR Panel Cartridge can detect common antimicrobial-resistant genes from the bloodstream. However, additional resistant genes should be included in the panel to cover various highly prevalent antimicrobial-resistant genes based on geographical location.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Personal views expressed by the contributors in their articles are not necessarily those of the Journal of Associated Medical Sciences, Faculty of Associated Medical Sciences, Chiang Mai University.
References
Jantarabenjakul W, Anugulruengkitt S, Puthanakit T, Pancharoen C. Drug-resistant bacteria: Drug-resistant organisms in pediatrics: Diagnosis and treatment. 1st Bangkok: Active Print; 2019: 288 [in Thai].
Global Database for Tracking Antimicrobial Resistance (AMR) Country Self-assessment Survey (TrACSS). Available from: https://amrcountryprogress.org/#/ download-responses.
Tancharoen L, Pairattanakorn P, Thamlikitkul V, Angkasekwinai N. Epidemiology and burden of sepsis at Thailand’s largest university-based National Tertiary Referral Center during 2019. Antibiotics. 2022 Jul 5;11(7):899.
Phumart P, Phodha T, Thamlikitkul V, Riewpaiboon A, Prakongsai P, Limwattananon S. Health and economic impacts of antimicrobial resistant infections in Thailand: A preliminary study. J Health Syst Res. 2012; 6(3): 352-60 (in Thai).
Indrawattana N, Vanaporn M. Nosocomial infection. J Med Health Sci. 2015; 22(1): 81-92.
Thawornwan U, Kongsanan P, Chaisukda S, Eampokalap B. Incidence of AmpC β-lactamase producing bacteria isolated in Bamrasnaradura Infectious Disease Institute during July-September 2007. J Bamrasnaradura Infect Dis Inst. 2007; 2(1): 32-9 (in Thai).
Wongut-sa P. Extended-spectrum β-Lactamases producing in Escherichia coli and Klebsiella pneumoniae isolated from specimens of patients in Ratchaburi Hospital (2009-2013). J Med Public Health Region 4. 2013; 17(3): 222-9 (in Thai).
Shanthi M, Sekar U, Arunagiri K, Sekar B. Detection of Amp C genes encoding for beta-lactamases in Escherichia coli and Klebsiella pneumoniae. Indian J Med Microbiol. 2012; 30(3): 290-5. doi: 10.4103/ 0255-0857.99489.
Reygaert WC. An overview of the antimicrobial resistance mechanisms of bacteria. AIMS Microbiol. 2018; 4(3): 482-501. doi: 10.3934/microbiol.2018. 3.482.
Rabaan AA, Eljaaly K, Alhumaid S, Albayat H, AlAdsani W Sabour AA, et al. An overview on phenotypic and genotypic characterisation of carbapenemresistant Enterobacterales. Medicina. 2022; 58(11): 1675. doi: 10.3390/medicina58111675.
Gharaibeh MH, Shatnawi SQ. An overview of colistin resistance, mobilized colistin resistance genes dissemination, global responses, and the alternatives to colistin: A review. Vet World. 2019; 12(11): 1735-46. doi: 10.14202/vetworld.2019.1735-1746.
Doi Y, Wachino JI, Arakawa Y. Aminoglycoside Resistance: The emergence of acquired 16S Ribosomal RNA methyltransferases. Infect Dis Clin North Am. 2016; 30(2): 523-37. doi: 10.1016/j.idc.2016.02.011.
Banerjee R, Patel R. Molecular diagnostics for genotypic detection of antibiotic resistance: current landscape and future directions. JAC Antimicrob Resist. 2023; 5(1): dlad018. doi.org/10.1093/jacamr/ dlad018.
Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. 34th CLSI supplement M100. Wayne: CLSI; 2024.
European Committee on Antimicrobial Susceptibility Testing. Antimicrobial susceptibility testing. 2024 [cited 2024 Nov 4]. Available from: https://www.eucast. org/ast_of_bacteria.
European committee on antimicrobial susceptibility testing. La Société française de microbiologie (SFM) guidelines. 2024 [cited 2024 Nov 4]. Available from: https://www.eucast.org/.
Ruekit S, Srijan A, Serichantalergs O, Margulieus KR, Mc Gann P, Mills EG, et al. Molecular characterization of multidrug-resistant ESKAPEE pathogens from clinical samples in Chonburi, Thailand (2017-2018). BMC Infect Dis. 2022; 22(1): 695. doi: 10.1186/ s12879-022-07678-8.
Kiattimanaroj A. The development of a model of care for patients with sepsis, Wapipathum Hospital. Acad J Mahasarakham Provincial Public Health Office. 2021; 5(9): 27 (in Thai).
Maenchantrarath C. Detection of bacteria in blood using the BactecTMFX: Procedure manual. Sparks: Becton Dickinson and Company; 2020.
Sun J, Zhao S, Wei C, Liu J, Chen J, Xing L, et al. A diagnostic test of real-time PCR detection in the diagnosis of clinical bloodstream infection. Ann Palliat Med. 2022; 11(10): 3224-30. doi: 10.21037/apm-22- 1071.
Trung NT, Thau NZ, Bang MH, Song LH. PCR-based sepsis@Quick test is superior in comparison with blood culture for identification of sepsis-causative pathogens. Sci Rep. 2019; 9(1): 13663. doi: 10.1038/ s41598-019-50150-y.
Department of Disease Control. Thailand’s integrated antimicrobial resistance surveillance with One Health Approach guideline. Nonthaburi: Ministry of Public Health; 2019: 112.
Roschanski N, Fischer J, Guerra B, Roesler U. Development of a multiplex real-time PCR for the rapid detection of the predominant beta-lactamase genes CTX-M, SHV, TEM and CIT-Type AmpCs in Enterobacteriaceae. PLoS ONE. 2014; 9(7): e100956. doi.org/10.1371/journal.pone.0100956.
Thummeepak R, Kongthai P, Leungtongkam U, Sitthisak S. Distribution of virulence genes involved in biofilm formation in multi-drug resistant Acinetobacter baumannii clinical isolates. Int Microbiol. 2016; 19(2): 121-9. doi: 10.2436/20.1501.01.270.
Juntanawiwat P, Thunyaharn S, Visawapoka U, Samosornsuk W, Samosornsuk S. The prevalence of OXA-type genes in carbapenem-resistant Acinetobacter baumannii isolated from critically ill patients in the intensive care unit of Phramongkutklao Hospital. J Med Tech Phys Ther. 2016; 28(2): 120-8.
Wongut-sa P. Khumdee P, Samosornsuk W Yansombat J, Samosornsuk S. Prevalence of colistin resistance among third-generation cephalosporins- or carbapenemresistant enterobacteriaceae Isolated from clinical specimens in Ratchaburi Hospital, Thailand. J Med Tech Assoc Thailand. 2020; 48(1): 60 (in Thai).
Ku YH, Lee MF, Chuang YC, Yu WL. Detection of plasmid- mediated β-lactamase genes and the emergence of a novel AmpC (CMH-1) in Enterobacter cloacae at a Medical Center in Southern Taiwan. J Clin Med. 2019; 8(1): 8. doi: 10.3390/jcm8010008.
Department of Medical Services, Health Systems Research Institute. EE-AMR Tool Thailand. 2021 [cited 2024 Nov 4]. Available from: https://kb.hsri.or.th/ dspace/handle/11228/5684.
Swe-Han K, Pillay M. Amikacin-resistant Acinetobacter species mediated by the aphA6 gene associated with clinical outcome at an academic complex hospital in KwaZulu-Natal Province. S Afr Med J. 2020; 110(1): 49-54. doi.org/10.7196/samj.2020.v110i1.14045.
Abouelfetouh A, Torky AS, Aboulmagd E. Phenotypic and genotypic characterization of carbapenemresistant Acinetobacter baumannii isolates from Egypt. Antimicrob Resist Infect Control. 2019 Dec;8(1):185. doi.org/10.1186/s13756-019-0611-6.
Ramachandran G. Gram-positive and gram-negative bacterial toxins in sepsis. Virulence. 2014; 5(1): 213-8. doi: 10.4161/viru.27024.
Wang Q, Li X, Tang W, Guan X, Xiong Z, Zhu Y, et al. Differential Gene Sets Profiling in GramNegative and Gram-Positive Sepsis. Front Cell Infect Microbiol. 2022 Feb 9;12:1-11. doi.org/10.3389/ fcimb.2022.801232.
Olaitan A, Morand S, Rolain JM. Mechanisms of polymyxin resistance: acquired and intrinsic resistance in bacteria. Front Microbiol. 2014; 5: 643. doi: 10.3389/ fmicb.2014.00643.