Blood Culture Bottles Research Articles

Performance evaluation of the Specific Reveal system for rapid antibiotic susceptibility testing from positive blood cultures containing Gram-negative pathogens.

Rapid antimicrobial drug administration is crucial for the efficient treatment of sepsis or septic shock, but empirical therapy is limited by the increasing prevalence of multidrug-resistant bacteria. Thus, rapid and reliable antimicrobial susceptibility testing (AST) is needed to start appropriate antimicrobial drug administration as quickly as possible. In the present study, we evaluated the performance of the Reveal rapid AST system. From February to April 2021, 102 positive blood culture bottles (BCBs) from hospitalized patients with bacteremia caused by Gram-negative bacteria were included in the study. All isolates were tested by the Reveal system directly from the positive BCBs in comparison to the DxM MicroScan WalkAway. Essential agreement (EA) and category agreement (CA) were high with 98.5% and 97.1%, respectively. We also determined the susceptibility of 10 highly resistant CDC & FDA AR strains in duplicate. Here, EA was 99.6% and CA 97.9%. The average time to result by Reveal was 5.4 h ± 1.2 h compared to an average of 16 h by DxM MicroScan WalkAway for clinical strains and 3.8 h ± 1.2 h for more resistant CDC & FDA AR strains. Susceptibility determination with the Reveal rapid AST system directly from positive BCBs is for the frequently represented bug-drug combinations a reliable and accurate approach, meeting the European ISO guideline for the performance of AST systems. Moreover, AST directly from blood cultures performed with the Reveal system saves time when compared to the conventional AST, as no subculturing is needed and time to result is very short.

Comparison of intra-assay and inter-assay reproducibility and positive detection times of two different (BacT/Alert 3D and Autobio BC) commercial blood culture systems.

In our study, we aimed to compare the performance of the BacT/Alert 3D (bioMerieux, France) system, which is currently used in our laboratory, and the Autobio BC (Autobio, China) system, which was newly introduced in our country, using standard and clinical isolates. Bacterial suspension was prepared by two technicians on the same day and three consecutive days from five different standard strains with 0.5 McFarland turbidity, then serial dilution to a final concentration was adjusted and was simultaneously inoculated in aerobic blood culture bottles. The bacterial concentration was measured by making a quantitative counting plate. The same procedure was also performed for 55 clinical isolates belonging to eleven species. After simulated bacteremia with standard and clinical isolates, the growth results were confirmed by inoculation from positive blood culture bottles onto solid medium and identification was made in the next day with MALDI-TOF MS (bioMérieux). In each study, sterile saline and blood was inoculated into the bottles as a negative control to check contamination. Intra-assay and inter-assay reproducibility of recovery rates and detection times of standard strains; recovery rates and detection times of clinical isolates were compared for both systems. Recovery rates were 100% in both systems, and when positive detection times were compared, it was found that there was no difference between the two devices in clinical isolates (p:0.262) but that Autobio BC gave significantly (p<0.001) earlier results in standard strains. In our simulated bloodstream infection study, Autobio BC was found to be comparable with BacT/Alert 3D, both recovery rates and growth detection time performance were found to be very good, and it can be used in routine microbiology laboratories.

Evaluation of the QIAstat-Dx BCID GN and GPF kits for direct identification and antimicrobial resistance prediction from blood culture bottles.

This is one of the first evaluations of the QIAstat BCID kit and demonstrates high levels of correlation for both identification and antimicrobial resistance prediction.

Rapid implementation of a clinical decision-support workflow during the national blood culture bottle shortage.

Rapid implementation of a clinical decision-support workflow during the national blood culture bottle shortage.

Culturomics: A promising approach for exploring bacterial diversity in natural fermented milk

Natural fermented milk is an abundant source of microbial resources, some of which possess probiotic properties that can help prevent and treat human diseases. However, many potential probiotics have not yet been isolated using traditional pure-culture methods. Culturomics, a relatively novel approach, can be primarily employed to cultivate and identify previously unknown and uncultured bacteria. Nevertheless, their application in natural fermented milk remains limited. Therefore, the primary objective of this study was to evaluate the bacterial diversity in natural fermented milk and explore its microbial potential. For this purpose, we collected three natural fermented milk samples from Inner Mongolia and subjected them to enrichment culture in blood culture bottles for 14 d. Bacterial isolates were obtained from the samples at 0, 3, 7, and 14 days using two culture temperatures and four media, and full-length 16S rRNA gene sequencing was performed for each sample at each time point. The results revealed that 488 strains were isolated from three samples belonging to 3 phyla, 11 genera, and 27 species. The method using two culture temperatures (20 °C and 30 °C) and two media (MRS and RCM + Vb) yielded most of the isolates after 3, 7, and 14 days of enrichment culture. Furthermore, full-length 16S rRNA gene sequencing confirmed that this method significantly increased bacterial diversity in natural fermented milk. In conclusion, the culturomics method demonstrates great potential for the isolation of a greater number of bacterial species and is advantageous for further exploration of the diversity of culturable microorganisms in natural fermented milk.

Clinical Practice: Estimating the Breakpoints for EUCAST Fast Antimicrobial Susceptibility Testing Using Flagged BacT/Alert Blood Culture Bottles

&amp;lt;i&amp;gt;Introduction&amp;lt;/i&amp;gt;: The escalating prevalence of multidrug resistance is a global threat to human health particularly in critically ill patients with bloodstream infections (BSIs). Delay in the administration of the appropriate antimicrobial treatment is associated with higher mortality rates and adverse consequences. This study attempted to estimate the rapid antimicrobial susceptibility testing (RAST) breakpoints directly from flagged BacT/Alert blood culture bottles in clinical practice. &amp;lt;i&amp;gt;Material &amp; Methods&amp;lt;/i&amp;gt;: A descriptive, cross-sectional study conducted at a tertiary care hospital in Delhi over a period of two months. The RAST was performed directly from the clinical samples for blood cultures received in our laboratory in parallel with the routine antimicrobial testing as per standard CLSI guidelines. Blood cultures were routinely incubated in BacT/Alert 3D. The inhibition zones were read at 4, 6, 8 and 16-20 hour of incubation as per European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. The identification of the isolates was confirmed by Vitek-2 compact system. &amp;lt;i&amp;gt;Results&amp;lt;/i&amp;gt;: In our study, the area of technical uncertainty (ATU) percentage was initially high at 4 hours but decreased significantly in later incubation periods. At 4 hours, none of the &amp;lt;i&amp;gt;S. aureus&amp;lt;/i&amp;gt; isolates showed &amp;gt;90% categorical agreement (CA) for any antimicrobial tested. However, clindamycin achieved the highest CA (100%) at 6 hours and 90% thereafter, with no very major errors (VME) or major error (ME). Cefoxitin required 8 hours to reach &amp;gt;90% CA, with no VME observed at any time point, but up to 75% ME at 8 hours. At 4 hours, most antimicrobials had high (&amp;gt;1.5%) rates of VME among &amp;lt;i&amp;gt;Enterobacteriales&amp;lt;/i&amp;gt;. By 6 hours, only Meropenem and Gentamicin had &amp;gt;90% CA, with no VME observed for other antibiotics. &amp;lt;i&amp;gt;Conclusion&amp;lt;/i&amp;gt;: The RAST method is relatively easy to implement in clinical microbiology labs, offering cost-effectiveness, simplicity, and rapid results, especially in resource-limited settings. However, reporting RAST results can be complex due to potential challenges with CA, VME, and ME, particularly in the initial hours of incubation and within the ATU.

Improving the microbiological diagnosis of fracture-related infection and prosthetic joint infection through culturing sonication fluid in Bactec blood culture bottles.

Sonication of surgically removed implants appears to optimize the microbiological diagnosis in orthopedic implant-associated infections (OIAI). However, reports of infection with negative cultures can still reach high rates. A study evaluating the inoculation of sonication fluid into blood culture bottles (SFBCB) in patients with fracture-related infection (FRI) and prosthetic joint infection (PJI) is necessary. This study compared the accuracy SFBCB over the conventional sonication fluid cultures (CSFC) and tissue culture (TC). Consecutive patients who underwent implant removal surgeries due to any reason had their implants sonicated according to standardized method. Definitions of PJI and FRI were based upon criteria by European Bone and Joint Infection Society (EBJIS) and Metsemakers, respectively. Between three to five intraoperative tissue samples were processed. The implant`s sonication fluid was seeded onto sheep blood agar, chocolate agar, thioglycolate broth and on tryptic soy broth for CSFC, while was also inoculated into blood culture bottles and incubated in the automated system during 5 days for SFBCB. Overall, 74 patients were analyzed, of which 57 with OIAI (48 FRI and 09 PJI) and 17 aseptic failures (03 arthroplasties and 14 osteosynthesis). Interestingly, SFBCB demonstrated significantly higher sensitivity compared to CSFC (96.5% [95% CI, 88-100] vs. 78.9% [95% CI, 66-89], p = 0.004), and to TC (96.5% [95% CI, 88-100], vs. 57.9% [95% CI, 44-71], p < 0.001), whereas there were no significant differences in specificity between the three methods. In comparison to CSFC and TC, SFBCB improved sensitivity for diagnosing OIAI without compromising specificity.

Accuracy of various matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based rapid identification methods—As a tool to augment diagnostic stewardship in blood culture laboratory, South India

BackgroundBlood stream infection is a medical emergency associated with high morbidity and mortality. Prompt identification of bloodstream infection-causing microorganisms directly from positive blood culture will significantly enhance patient care by reducing the turnaround time of pathogen recognition. MethodsA total of 256 freshly flagged positive blood culture bottles were subjected to Gram staining. Direct MALDI-TOF MS analysis was performed following sample preparation techniques such as lysis centrifugation, lysis filtration and VITEK® MS BC kit to directly identify microorganisms from positive blood cultures. Along with these short-term incubation methods of Choco spot and minute colony(8–10h) were also performed. All those positive bottles were identified by the routine (reference) laboratory method. Results177 isolates (69.14 %) were correctly identified by Lysis centrifugation, 163 isolates (63.67 %) were correctly identified by Lysis filtration, 206 isolates (80.47 %) were correctly identified by Choco spot,250 isolates (97.65 %) were correctly identified from minute colony (8–10h) of incubation. Of 162 isolates,115 isolates (70.99 %) were correctly identified by VITEK® MS Blood culture kit, (BioMérieux). VITEK® MS BC kit method revealed higher agreement with the kappa value of 0.697 than lysis centrifugation (0.672) followed by lysis filtration (0.611). ConclusionsIn house method of lysis centrifugation is found to be equivalent to VITEK® MS BC kit method and superior to lysis filtration method in correct direct identification of bacteria from positive blood cultures by MALDI-TOF MS analysis. As lysis centrifugation requires only 10 min of processing time as compared to overnight incubation, thus it offers a less expensive substitute for the VITEK® MS BC kit in the clinical laboratory. As a consequence of this study, we have implemented direct MALDI-TOF-based identification from positive BCs in our daily routine diagnostic management.

Distribution and antifungal susceptibility profiles of Candida species isolated from candidemia patients admitted to Egyptian tertiary hospitals: a cross-sectional study

BackgroundCandidemia is a widespread threat that can lead to significant complications in healthcare settings.ObjectivesOur study aimed to identify isolates of Candida isolated from blood culture bottles of patients with candidemia and assess their antifungal susceptibility profiles.MethodsWe conducted a cross-sectional study at Cairo University tertiary care hospitals over 16 months including 90 patients. Candida isolates were collected from blood culture bottles, and identified using MALDI-TOF MS technology of VITEK MS PRIME (bioMérieux) with the corresponding database VITEK IVD Database 3.2. followed by antifungal susceptibility testing using VITEK 2 Compact system.ResultsCandida albicans was the most common species isolated from both pediatric and adult patients with percentages of 47.3% and 36.4% respectively, followed by Candida parapsilosis with percentages of 32.6% and 25.0% respectively. Voriconazole showed the highest antifungal activity at 90.9% of isolates in adults and 95.7% in pediatrics, followed by caspofungin and micafungin. The mean hospital stays for adults ranged from 8 to 30 days and from 10 to 42 days in the pediatric group.ConclusionsC. albicans remains the predominant species isolated from both pediatric and adult candidemia patients, despite a notable increase in other species. C. tropicalis and C. parapsilosis are considered the most common non-albicans Candida (NAC) species. The rise in Candida species other than albicans highlights the urgent need for effective antifungal stewardship programs. Voriconazole exhibited the higher antifungal activity followed by caspofungin and micafungin.

Blood culture bottles meet the operating room: enhancing the diagnostic accuracy of infectious spondylitis through open microsurgical biopsy and intraoperative inoculation.

Infectious spondylitis is caused by hematogenous seeding or adjacent soft tissue infection. No study has provided evidence that incubating biopsy specimens in blood culture bottles could enhance detection rates, nor has any study compared this method with conventional culture techniques. We aimed to assess the diagnostic yield of open microsurgical biopsies for infectious spondylitis and the efficacy of various culture media in the presence and absence of pre-biopsy antibiotic therapy. This retrospective study, which was conducted at a university-affiliated teaching hospital in Korea, enrolled 165 adult patients with suspected infectious spondylitis between February 2014 and September 2020. The diagnostic yield of open biopsy was compared among three culture media, namely, blood culture bottles, swab culture using transport media, and tissue culture using plain tubes, while considering preoperative antibiotic exposure. Causative bacteria were identified in 84.2% of all cases. Blood culture bottles had the highest positivity rate (83.5%), followed by swab cultures (64.4%) and tissue cultures (44.9%). The differences in positivity rates were significant (P < 0.001). Preoperative antibiotic therapy reduced detection rates across all media, particularly in tissue cultures. We established the high diagnostic yield of open microsurgical biopsy using blood culture bottles, suggesting that pre-biopsy antibiotic therapy significantly affects bacterial detection, thereby underscoring the importance of culture medium selection in the diagnosis of infectious spondylitis.

Impact of direct identification of bacteria in blood culture–positive specimens by matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry on physician selection of antimicrobial therapy

BackgroundRapid identification of the causative organism in blood stream infections is essential for early initiation of appropriate antimicrobial therapy. Direct identification of bacteria in positive blood culture bottles using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry is a promising application. A variety of direct identification methods have been reported; however, few studies have evaluated the impact of these methods on physician decision making regarding antimicrobial therapy. MethodsWe developed a simple method for direct bacterial identification and applied it to daily clinical practice to investigate the impact of direct identification of bacteria in positive blood culture bottles on physicians’ choice of antimicrobial agents for treatment. ResultsFrom January 2016 to December 2022, we attempted direct identification in 98 cases and successfully acquired identification results in 88 cases. In three cases, no empiric antimicrobial agents were initiated at the time of venipuncture for blood culture but later initiated based on the direct identification results. In the remaining 85 cases, empiric antimicrobial therapy was initiated at the time blood cultures were performed, and in 29 cases, empiric antimicrobial therapy was changed after direct identification. In 17 of these 29 cases, the antimicrobial therapy was changed based on the direct identification of bacterial genus/species, resulting in a change to an effective antimicrobial therapy before the antimicrobial susceptibility testing results were available. ConclusionsDirect identification of bacteria from positive blood culture bottles could contribute to earlier selection of or changes to antimicrobial therapies by attending physicians.

Pseudo-outbreak of Bacillaceae species associated with poor compliance with blood culture collection recommendations.

This study describes a pseudo-outbreak of Bacillaceae spp. bloodstream infections that spanned five months starting in May 2023 and the infection prevention measures implemented to control it. This retrospective study was conducted at a tertiary infectious disease hospital in Bucharest, Romania. An observational audit of the blood culture collection practice in our hospital was performed, and the materials used during blood culture collection were sampled. Bacterial colonies were identified using MALDI Biotyper. The Bacillaceae blood culture positivity rates in the previous four years were compared using the Kruskal‒Wallis rank test. Bacillaceae spp.-positive blood cultures were recovered from 60 patients over a five-month period. In the case of 58 patients, Bacillaceae spp.-positive blood cultures were considered contaminated. Two patients were treated for Bacillus spp. bacteraemia. The audit revealed significant variation during the preparation of the venipuncture site step and the use of nonsterile medical cotton wool. Medical cotton wool contaminated with species of Bacillaceae was found in 10 out of 12 wards. The control measures included repeated training on the blood culture collection procedure and the removal of Bacillaceae spp.-contaminated cotton wool. The pseudo-outbreak was caused by the unjustified use of medical cotton wool for disinfection of the skin and blood culture bottle septums. The investigation of this pseudo-outbreak highlighted a gap in blood culture collection practices at our facility and thus allowed for its improvement.

A comparative study of a rapid phenotypic antimicrobial susceptibility testing system directly from positive blood cultures to the disk diffusion and VITEK 2 methods

BackgroundSepsis is a life-threatening condition that impacts 49 million people annually and causes 11 million deaths worldwide. Surviving bloodstream infections (BSIs) depends on the rapid administration of effective antimicrobial treatment, underscoring a need for rapid antimicrobial susceptibility testing (AST). AimTo evaluate the performance of Quantamatrix's dRAST v2.5 system (Seoul, South Korea) for AST directly from positive blood cultures as compared to the Disk-Diffusion (DD) and VITEK 2 methods. MethodsThe study included 191 positive blood cultures from clinical samples and spiked blood culture bottles. Following Gram staining and species-level identification, AST was performed by VITEK 2 and standard DD methods using CLSI (2021) interpretation. ResultsdRAST demonstrated very good AST performance for a Gram-negative isolate, and good performance for Gram-positive isolates, meeting CLSI criteria for the acceptance of a new method. Antimicrobials that were not considered verified compared to VITEK 2 and DD were cefazolin, ceftazidime, meropenem, and trimethoprim/sulfamethoxazole for Gram-negatives and clindamycin, erythromycin, penicillin, and oxacillin for Gram-positives. dRAST ESBL detection results were strongly correlated with the ESBL phenotypes obtained with other methods. Additional resistance mechanisms were in concordance with traditional tests. ConclusionsdRAST demonstrated good AST performance, meeting CLSI criteria for most relevant antibiotics. dRAST was associated with a significant reduction in time-to-results, labor, and the subjectivity of result analyses, making it a valuable addition to efforts supporting the treatment of patients with bacteremia.AST (antimicrobial susceptibility test), blood culture, dRAST, rapid methods, sepsis, turnaround time (TAT).

Comparison of Zybio Kit and saponin in-house method in rapid identification of bacteria from positive blood cultures by EXS2600 matrix-assisted laser desorption ionization time-of-flightmass spectrometry system.

We evaluated the performance of Zybio EXS2600 matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) (Zybio Inc., Chongqing, China) for the identification of bacteria from positive blood culture (BC) bottles using Blood Culture Positive Sample Pretreatment Kit (Zybio Inc., Chongqing, China) in comparison to an in-house saponin method. Following a positive signal by the BACTEC™ FX system, confirmation of identification was achieved using subcultured growing biomass used for MALDI-TOF MS analysis. A total of 94 positive BC bottles with 97 bacterial isolates were analyzed. The overall identification rates at the genus and species levels for the saponin method were 89.7% (87/97) and 74.2% (72/97), respectively. With the Zybio Kit, 88.7% (86/97) and 80.4% (78/97) of microorganisms were correctly identified to the genus and species levels, respectively. The saponin method identified 65.3% (32/49) of Gram-positive bacteria at the species level, whereas the Zybio Kit achieved a higher species-level identification rate of 79.6% (39/49) (p= 0.1153). The saponin method with additional on-plate formic acid extraction showed a significantly higher overall identification rate in comparison to the saponin method without that step for both genus (87.6% [85/97] vs. 70.1% [68/97], p= 0.0029) and species level (70.1% [68/97] vs. 46.4% [45/97], p= 0.0008). Identification rates of Gram-negative bacteria showed a higher identification rate, however, not statistically significant with additional Zybio Kit protocol step on both genus (85.4% [41/48] vs. 81.3% [39/48], p= 0.5858) and species level (77.1% [37/48] vs. 75% [36/48], p= 0.8120). Zybio Kit could offer an advantage in species-level identification, particularly for Gram-positive bacteria. The inclusion of on-plate formic acid extraction in the saponin method notably enhanced identification at both genus and species levels for Gram-positive bacteria. The extended protocol provided by the Zybio Kit could potentially offer an advantage in the identification of Gram-negative bacteria at both genus and species levels. Enhancements to the Zybio EXS2600 MALDI-TOF instrument software database are necessary.

Evaluation of the Reveal® AST (SPECIFIC) for Antimicrobial Susceptibility Testing from Positive Blood Culture Spiked with Carbapenem-Resistant Isolates.

As bloodstream infections and associated septic shock are common causes of mortality in hospitals, rapid antibiotic susceptibility testing (AST) performed directly on positive blood cultures is needed to implement an efficient therapy in clinical settings. We evaluated the Reveal® rapid AST system on a collection of 197 fully characterized carbapenem-resistant Enterobacterales, including 177 carbapenemase producers (CPE) spiked in blood culture bottles. The clinical categorization based on the Minimal Inhibitory Concentration (MIC) determination of eighteen antimicrobial molecules was compared to the clinical categorization based on the disk diffusion assay as a reference. The Reveal AST system provided results within a mean time to result of 5 h. Overall, the categorical agreement (CA) between the two techniques was 94.1%. The rates of very major errors (VMEs), major errors (MEs) and minor errors (mEs) were 3.8%, 3.7% and 5.6%, respectively. Imipenem was the antimicrobial with the lowest CA rate (78.7%), with rates of 15% VMEs and 10.7% MEs, but the performances were better when considering only the non-CPE category (CA of 89%). On this resistant collection of Enterobacterales with numerous acquired β-lactamases, the Specific Reveal assay proved to be useful for a rapid determination of AST compatible with a quick adaptation of the patient's antimicrobial treatment.

Evaluation of Direct Antimicrobial Susceptibility Testing of Gram-Negative Bacilli and Staphylococcus aureus from Positive Pediatric Blood Culture Bottles Using BD Phoenix M50.

Bloodstream infections (BSIs) are life-threatening infections for which a timely initiation of appropriate antimicrobial therapy is critical. Antibiotic susceptibility testing (AST) directly performed on positive blood culture broths can help initiate targeted antibiotic therapy sooner than the standard AST performed on colonies isolated on solid media after overnight incubation. Faster antimicrobial susceptibility testing (AST) results can improve clinical outcomes, and reduce broad-spectrum antimicrobial consumption and healthcare-associated costs in sepsis. In this study, we evaluated the accuracy of a direct AST inoculation method on the BD Phoenix M50 system using serum separator tubes to harvest bacteria from positive pediatric blood culture bottles. Direct AST was performed on 132 monomicrobial pediatric blood culture bottles that were positive for Enterobacterales (65; 49.2%), Staphylococcus aureus (46; 34.8%), and non-fermenting Gram-negative bacilli (21; 16%). Overall, the categorical and essential agreements between the direct method and standard method were 99.6% and 99.8%, respectively. Very major, major, and minor error rates were 0.1%, 0.09%, and 0.20% respectively. Direct AST performed on pediatric blood culture bottles using BD Phoenix M50 can quickly provide accurate susceptibility information to guide antimicrobial therapy in patients with BSI.

Rapid implementation of blood culture stewardship: institutional response to an acute national blood culture bottle shortage.

We describe our approach to addressing a nation-wide supply issue for blood culture bottles. Aerobic blood culture bottles received from our distributor July 1-15, 2024 was <1% of typical usage. Through education and ordering restrictions blood culture designed to minimize risk, orders were reduced by 49% over a one-week period.

Investigation of different methods in rapid microbial identification directly from positive blood culture bottles by MALDI-TOF MS.

Sepsis is a life-threatening condition, and rapid and accurate identification of the causative microorganisms from blood cultures is crucial for timely and effective treatment. Although there are many studies on direct identification from blood cultures with MALDI-TOF MS, further standardization is still needed. In our study, we analyzed the performance of three different preparation methods and compared by using two analysis modules of the Bruker Biotyper MALDI-TOF MS for direct identification of bacteria from numerous positive blood culture bottles. The literature reports a limited number of studies that compare different preparation methods for direct blood culture identification, processing a large number of blood samples concurrently and evaluating the same samples as in our study. Moreover, although SDS is used very frequently in medical laboratories, there are few studies on direct identification from blood culture bottles. In our study, the highest correct identification rate was observed with the SDS method.

Microbiological performance and adherence in blood culture protocols: The role of a second anaerobic bottle

BackgroundBacteremia represents high rates of morbidity and mortality, especially in developing countries, highlighting the need for a diagnostic method that allows prompt and appropriate patient treatment. This study compared microbiological performance and adherence of two blood culture protocols for the diagnosis of bacteremia. MethodsQuasi-experimental study conducted between June 2022 and February 2023. Two blood culture protocols were evaluated. Protocol 1 included two aerobic bottles and one anaerobic bottle. Protocol 2 included two aerobic and two anaerobic bottles. Protocols were analyzed in three phases: evaluation of protocol 1 (Phase 1); evaluation of protocol 1 plus educational activities for healthcare staff (Phase 2) and evaluation of protocol 2 (Phase 3). Results342 patients and 1155 blood culture bottles (732 aerobic and 423 anaerobic) were included. Positivity was 17.6 %, 22.8 % and 19.4 % in phases 1, 2 and 3, respectively. Among patients with bacteremia, 84.5 % had positive anaerobic bottles, with 9.9 % showing growth only in this bottle. The contamination rates were 1.9 %, 0.3 %, and 0.8 % for each phase, mainly in aerobic bottles. Median positivity time was 11 h for both bottes aerobic and anaerobic. Overall nursing adherence increased from 13.1 % in Phase 1, 25.9 % in Phase 2, and 28.1 % in Phase 3 (p = 0.009). ConclusionsThe findings indicate that adding a second anaerobic bottle does not enhance blood culture positivity. Rather than increasing bottle quantity, staff training might be a more effective approach to optimize results.

Gas Chromatography-Ion Mobility Spectrometry Reveals Acetoin as a Biomarker for Carbapenemase-Producing Klebsiella pneumoniae.

BACKGROUND This study aimed to detect the volatile organic compound (VOC), 3-hydroxy-2-butanone (acetoin) using gas chromatography-ion mobility spectrometry (GC-IMS) in antimicrobial-resistant Klebsiella pneumoniae (K. pneumoniae) carbapenemase (KPC)-producing bacteria. MATERIAL AND METHODS Using stromal fluid of blood culture bottles (BacT/ALERT® SA) as the medium, 3-hydroxy-2-butanone (acetoin) released by K. pneumoniae during growth was detected using GC-IMS. The impact of imipenem (IPM) and carbapenemase inhibitors [avibactam sodium or pyridine-2,6-dicarboxylic acid (DPA)] on the emission of 3-hydroxy-2-butanone (acetoin) from various carbapenemase-producing K. pneumoniae was further investigated. Subsequently, VOCal software was used to generate a pseudo-3D plot of 3-hydroxy-2-butanone (acetoin), and the relative peak volumes were exported for data analysis. Standard strains served as references, and the findings were validated with clinical isolates. RESULTS The pattern of temporal changes in the 3-hydroxy-2-butanone (acetoin) release from K. pneumoniae in the absence of IPM was consistent with the growth curve. After the IPM addition, carbapenemase-positive strains released significantly higher contents of 3-hydroxy-2-butanone (acetoin) than carbapenemase-negative strains at the late exponential growth phase (T2). Notably, adding avibactam sodium significantly decreased the 3-hydroxy-2-butanone (acetoin) content released from the class A carbapenemase-producing strains as compared to the absence of the carbapenemase inhibitor. Conversely, adding DPA significantly decreased the 3-hydroxy-2-butanone (acetoin) content released from the class B carbapenemase-producing strains (both standard and clinical strains, all P<0.05). CONCLUSIONS This study demonstrated the potential of 3-hydroxy-2-butanone (acetoin) as a VOC biomarker for detecting carbapenemase-producing K. pneumoniae, as revealed by GC-IMS analysis.