Abstract
Aim: In patients with bloodstream infection; identifying the causative agent in a short time and applying appropriate antimicrobial therapy is very important in terms of reducing morbidity and mortality. In this study, it was aimed to evaluate a new method developed for direct identification and antibiotic susceptibility testing from blood culture.
Materials and Methods: Bacterial pellet was obtained from blood culture samples that showed positive signals in the BacT/Alert 3D system by washing and centrifugation. Identification was made from this pellet using Vitek MS, followed by direct antibiotic susceptibility testing in the VITEK 2 automated system. The results were compared with the standard method.
Results: Identification was performed on 80 blood culture samples. In the direct identification process, 73 samples were identified, of which 72 (90%) resulted in agreement with the standard method. Direct antibiotic susceptibility tests were found to be compatible with a rate of 97.9%. Among the evaluated 635 antibiotic susceptibility results; it was observed that 10 of them had major errors and 3 of them had minor errors.
Conclusions: Using the procedures applied in the study from blood culture samples, without the need for costly reagents or long processing time, it has been observed that identification and antibiotic susceptibility results can be obtained within 24 hours, similar to standard results.
References
- Kadri SS, Lai YL, Warner S, Strich JR, Babiker A, Ricotta EE, et al. Inappropriate empirical antibiotic therapy for bloodstream infections based on discordant in-vitro susceptibilities: a retrospective cohort analysis of prevalence, predictors, and mortality risk in US hospitals. Lancet Infect Dis. 2021 Feb; 21 (2): 241-251. doi: 10.1016/S1473-3099 (20) 30477-1. Epub 2020 Sep 8. PMID: 32916100
- Kumar A, Roberts D,Wood KE, Light B, Parrillo JE, Sharma S, et al. Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 2006; 34: 1589–96.
- Sauer S, Kliem M. Mass spectrometry tools for the classification and identification of bacteria. Nat Rev Microbiol 2010; 8: 74–82.
- Ferroni A, Suarez S, Beretti JL, Dauphin B, Bille E, Meyer J, et al. Real-time identification of bacteria and Candida species in positive blood culture broths bymatrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2010; 48: 1542–8.
- Buchan BW, Riebe KM, Ledeboer NA. Comparison of the MALDI Biotyper system using Sepsityper specimen processing to routine microbiological methods for identification of bacteria from positive blood culture bottles. J Clin Microbiol 2012; 50: 346–52.
- Azrad M, Keness Y, Nitzan O, Pastukh, Tkhawkho, Freidus V, et al. Cheap and rapid in-house method for direct identification of positive blood cultures by MALDI-TOF MS technology. BMC Infect Dis. 2019 Jan 18; 19 (1): 72. doi: 10.1186/s12879-019-3709-9.
- Lin JF, Ge MC, Liu TP, Chang SC, Lu JJ. A simple method for rapid microbial identification from positive monomicrobial blood culture bottles through matrix-assisted laser desorption ionization time-of-flight mass spectrometry. J Microbiol Immunol Infect. 2018 Oct; 51 (5): 659-665. doi: 10.1016/j.jmii.2017.03.005. Epub 2017 Jun 30.
- Hannah Tanner, Jason T. Evans, Savita Gossain, Abid Hussain. Evaluation of three sample preparation methods for the direct identification of bacteria in positive blood cultures by MALDI‑TOF. BMC Res Notes. 2017; 10: 48. doi: 10.1186/s13104-016-2366-y.
- Pan H, Li W, Li R, Li Y, Zhang Y, Sun E. Simple Sample Preparation Method for Direct Microbial Identification and Susceptibility Testing From Positive Blood Cultures. Front Microbiol. 2018 Mar 20; 9: 481. doi: 10.3389/fmicb.2018.00481.
- Maelegheer K., Nulens E. Same-day identification and antibiotic susceptibility testing on positive blood cultures: a simple and inexpensive procedure. Eur J Clin Microbiol Infect Dis (2017) 36: 681–687 DOI 10.1007/s10096-016-2849-8.
- Rodríguez-Sánchez B, Sánchez-Carrillo C, Ruiz A, Marín M, Cercenado E, Rodríguez-Créixems M, Bouza E. Direct identification of pathogens from positive blood cultures using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. Clin Microbiol Infect. 2014 Jul; 20 (7): O421-7. doi: 10.1111/1469-0691.12455.
- Fothergill A, Kasinathan V, Hyman J, Walsh J, Drake T, Wang YF. Rapid identification of bacteria and yeasts from positive BacT/ALERT blood culture bottles by using a lysis-filtration method and MALDI-TOF mass spectrum analysis with SARAMIS database. J Clin Microbiol 2013: 51: 805–9.
- Clerc O, Prod’hom G, Vogne C, Bizzini A, Calandra T, Greub G. Impact of matrix-assisted laser desorption ionization time-of-flight mass spectrometry on the clinical management of patients with gramnegative bacteremia: a prospective observational study. Clin Infect Dis 2013; 56: 1101–7.
- Höring S, Massarani A, Löffler B, Rödel J. Rapid antibiotic susceptibility testing in blood culture diagnostics performed by direct inoculation using the VITEK®-2 and BD Phoenix™ platforms. European Journal of Clinical Microbiology & Infectious Diseases (2019) 38: 471 478. doi.org/10.1007/s10096-018-03445-3.
- López-Pintor J, Navarro-San Francisco C, Sánchez-López J, García-Caballero A, Bobadilla R, Morosini M. Direct antimicrobial susceptibility testing from the blood culture pellet obtained for MALDI-TOF identification of Enterobacterales and Pseudomonas aeruginosa. European Journal of Clinical Microbiology & Infectious Diseases doi.org/10.1007/s10096-019-03498-y.
Abstract
Amaç: Kan dolaşımı enfeksiyonu olan hastalarda; etkenin kısa sürede tanımlanması ve uygun antimikrobiyal tedavi uygulanması, morbidite ve mortalitenin azaltılması bakımından oldukça önemlidir. Bu çalışmada, kan kültüründen doğrudan tanımlama ve antibiyotik duyarlılık testlerinin yapılması için geliştirilen yeni bir yöntemin değerlendirilmesi amaçlanmıştır.
Gereç ve Yöntem: BacT/Alert 3D sisteminde pozitif sinyal veren kan kültürü örneklerinden yıkama ve santrifüj işlemleri ile bakteriyel çökelti elde edildi. Bu çökeltiden Vitek MS kullanılarak tanımlama yapıldı, ardından VITEK 2 otomatize sisteminde doğrudan antibiyotik duyarlılık testi çalışıldı. Sonuçlar standart yöntem ile karşılaştırıldı.
Bulgular: Tanımlama işlemi 80 kan kültürü örneğinde gerçekleştirildi. Doğrudan tanımlama işleminde 73 örnek tanımlandı ve bunlardan 72’si (%90) standart yöntemle uyumlu olarak sonuçlandı. Doğrudan antibiyotik duyarlılık testlerinin %97,9 oranında uyumlu olduğu saptandı. Değerlendirilen 635 antibiyotik duyarlılık sonucu içinde; 10’unda büyük hata, 3’ünde küçük hata olduğu görüldü.
Sonuç: Kan kültürü örneklerinden çalışmada uygulanan prosedürler kullanılarak 24 saat içinde, maliyetli reaktifler ya da uzun işlem süresine gereksinim olmadan, standart uygulama sonuçlarına benzer bir şekilde tanımlama ve antibiyotik duyarlılık sonucu elde edilebileceği görülmüştür.
Keywords
Supporting Institution
EGE ÜNİVERSİTESİ TIP FAKÜLTESİ BAP
Thanks
Saygıdeğer hocam Prof. Dr. Şöhret Aydemir'e desteklerinden ötürü çok teşekkür ederim.
References
- Kadri SS, Lai YL, Warner S, Strich JR, Babiker A, Ricotta EE, et al. Inappropriate empirical antibiotic therapy for bloodstream infections based on discordant in-vitro susceptibilities: a retrospective cohort analysis of prevalence, predictors, and mortality risk in US hospitals. Lancet Infect Dis. 2021 Feb; 21 (2): 241-251. doi: 10.1016/S1473-3099 (20) 30477-1. Epub 2020 Sep 8. PMID: 32916100
- Kumar A, Roberts D,Wood KE, Light B, Parrillo JE, Sharma S, et al. Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 2006; 34: 1589–96.
- Sauer S, Kliem M. Mass spectrometry tools for the classification and identification of bacteria. Nat Rev Microbiol 2010; 8: 74–82.
- Ferroni A, Suarez S, Beretti JL, Dauphin B, Bille E, Meyer J, et al. Real-time identification of bacteria and Candida species in positive blood culture broths bymatrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2010; 48: 1542–8.
- Buchan BW, Riebe KM, Ledeboer NA. Comparison of the MALDI Biotyper system using Sepsityper specimen processing to routine microbiological methods for identification of bacteria from positive blood culture bottles. J Clin Microbiol 2012; 50: 346–52.
- Azrad M, Keness Y, Nitzan O, Pastukh, Tkhawkho, Freidus V, et al. Cheap and rapid in-house method for direct identification of positive blood cultures by MALDI-TOF MS technology. BMC Infect Dis. 2019 Jan 18; 19 (1): 72. doi: 10.1186/s12879-019-3709-9.
- Lin JF, Ge MC, Liu TP, Chang SC, Lu JJ. A simple method for rapid microbial identification from positive monomicrobial blood culture bottles through matrix-assisted laser desorption ionization time-of-flight mass spectrometry. J Microbiol Immunol Infect. 2018 Oct; 51 (5): 659-665. doi: 10.1016/j.jmii.2017.03.005. Epub 2017 Jun 30.
- Hannah Tanner, Jason T. Evans, Savita Gossain, Abid Hussain. Evaluation of three sample preparation methods for the direct identification of bacteria in positive blood cultures by MALDI‑TOF. BMC Res Notes. 2017; 10: 48. doi: 10.1186/s13104-016-2366-y.
- Pan H, Li W, Li R, Li Y, Zhang Y, Sun E. Simple Sample Preparation Method for Direct Microbial Identification and Susceptibility Testing From Positive Blood Cultures. Front Microbiol. 2018 Mar 20; 9: 481. doi: 10.3389/fmicb.2018.00481.
- Maelegheer K., Nulens E. Same-day identification and antibiotic susceptibility testing on positive blood cultures: a simple and inexpensive procedure. Eur J Clin Microbiol Infect Dis (2017) 36: 681–687 DOI 10.1007/s10096-016-2849-8.
- Rodríguez-Sánchez B, Sánchez-Carrillo C, Ruiz A, Marín M, Cercenado E, Rodríguez-Créixems M, Bouza E. Direct identification of pathogens from positive blood cultures using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. Clin Microbiol Infect. 2014 Jul; 20 (7): O421-7. doi: 10.1111/1469-0691.12455.
- Fothergill A, Kasinathan V, Hyman J, Walsh J, Drake T, Wang YF. Rapid identification of bacteria and yeasts from positive BacT/ALERT blood culture bottles by using a lysis-filtration method and MALDI-TOF mass spectrum analysis with SARAMIS database. J Clin Microbiol 2013: 51: 805–9.
- Clerc O, Prod’hom G, Vogne C, Bizzini A, Calandra T, Greub G. Impact of matrix-assisted laser desorption ionization time-of-flight mass spectrometry on the clinical management of patients with gramnegative bacteremia: a prospective observational study. Clin Infect Dis 2013; 56: 1101–7.
- Höring S, Massarani A, Löffler B, Rödel J. Rapid antibiotic susceptibility testing in blood culture diagnostics performed by direct inoculation using the VITEK®-2 and BD Phoenix™ platforms. European Journal of Clinical Microbiology & Infectious Diseases (2019) 38: 471 478. doi.org/10.1007/s10096-018-03445-3.
- López-Pintor J, Navarro-San Francisco C, Sánchez-López J, García-Caballero A, Bobadilla R, Morosini M. Direct antimicrobial susceptibility testing from the blood culture pellet obtained for MALDI-TOF identification of Enterobacterales and Pseudomonas aeruginosa. European Journal of Clinical Microbiology & Infectious Diseases doi.org/10.1007/s10096-019-03498-y.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Health Care Administration |
| Journal Section | Research Articles |
| Authors | |
| Publication Date | June 13, 2022 |
| Submission Date | September 13, 2021 |
| Published in Issue | Year 2022Volume: 61 Issue: 2 |
