Rapid Direct Identification of Microbial Pathogens and Antimicrobial Resistance Genes in Positive Blood Cultures Using a Fully Automated Multiplex PCR Assay.

This study assessed the performance of the BioFire Blood Culture Identification 2 (BCID2) panel in identifying microorganisms and antimicrobial resistance (AMR) profiles in positive blood cultures (BCs) and its influence on turnaround time (TAT) compared with conventional culture methods. We obtained 117 positive BCs, of these, 102 (87.2%) were correctly identified using BCID2. The discordance was due to off-panel pathogens detected by culture (n = 13), and additional pathogens identified by BCID2 (n = 2). On-panel pathogen concordance between the conventional culture and BCID2 methods was 98.1% (102/104). The conventional method detected 19 carbapenemase-producing organisms, 14 extended-spectrum beta-lactamase-producing Enterobacterales, 18 methicillin-resistant Staphylococcus spp., and four vancomycin-resistant Enterococcus faecium. BCID2 correctly predicted 53 (96.4%) of 55 phenotypic resistance patterns by detecting AMR genes. The TAT for BCID2 was significantly lower than that for the conventional method. BCID2 rapidly identifies pathogens and AMR genes in positive BCs.

Evaluation of an automated rapid phenotypic antimicrobial susceptibility testing (ASTar, Q-linea AB) applied directly on blood cultures bottles positive for Gram-negative pathogens.

We evaluated the performance of a new rapid phenotypic antimicrobial susceptibility test (ASTar; Q-linea AB) on Gram-negative bacilli, directly from positive blood cultures bottles. MIC values obtained by the routine reference method (Microscan, Beckman Coulter) were compared to the ones provided by the tested method (ASTar). ASTar demonstrated an overall essential agreement of 98% and a category agreement of 96.1%. The overall rate of major errors and very major errors was 2.5% and 3.3%, respectively. ASTar can represent a rapid, simple, and reliable method to speed up information about antimicrobial susceptibility of Gram-negative pathogens from positive blood culture bottles.

Changes in clinical parameters and inflammatory markers after blood culture collection facilitate early identification of positive culture in adult patients with COVID-19 and clinically suspected bloodstream infection.

OBJECTIVE: We explored whether changes in clinical parameters and inflammatory markers can facilitate early identification of positive blood culture in adult patients with COVID-19 and clinically suspected bloodstream infection (BSI). METHODS: This single-center retrospective study enrolled 20 adult patients with COVID-19 admitted to the intensive care unit who underwent blood culture for clinically suspected BSI (February 2020-November 2021). We divided patients into positive (Pos) and negative blood culture groups. Clinical parameters and inflammatory markers were obtained from medical records between blood culture collection and the first positive or negative result and compared between groups on different days. RESULTS: Patients in the positive culture group had significantly older age and higher D-dimer, immunoglobulin 6 (IL-6), and Sequential Organ Failure Assessment score as well as lower albumin (ALB). The area under the receiver operating characteristic curve (AUC) was 0.865 for IL-6, D-dimer and ALB on the first day after blood culture collection; the AUC was 0.979 for IL-6, IL-10, D-dimer, and C-reactive protein on the second day after blood culture collection. CONCLUSION: Changes in clinical parameters and inflammatory markers after blood culture collection may facilitate early identification of positive culture in adult patients with COVID-19 and clinically suspected BSI.

Pitfalls of a Multiplex PCR Panel for Identification of Bloodstream Pathogens.

BACKGROUND: We evaluated the diagnostic performance of the FilmArray Blood Culture Identification Panel (BCID; bioMerieux) for the detection of bloodstream pathogens. METHODS: From May to August 2022, up to 67 samples from positive blood cultures previously processed with BACTEC FX (BD) were collected and submitted to the BCID panel. BCID panel results were compared with traditional culture results. RESULTS: We tested 67 positive blood culture samples; 13 samples were from pediatric bottles of BACTEC Peds Plus/F media (BD). The overall sensitivity of the BCID panel was 89.9% (62/69; 95% CI, 80.2 - 95.3%). For blood-stream pathogens targeted by the BCID panel, sensitivity was 98.4% (62/63; 95% CI, 90.7 - > 99.9%). Interestingly, Proteus species were additionally detected in 6 samples from pediatric blood culture bottles. CONCLUSIONS: BCID demonstrated high clinical sensitivity for target pathogens, but positive findings for unexpected multiple targets or Proteus species require cautious interpretation to avoid false positives.

Rapid antibiotic susceptibility testing (RAST) of blood cultures in enterobacteria with inducible chromosomal AmpC-type ß-lactamase.

INTRODUCTION: Early and adequate treatment of bloodstream infections decreases patient morbidity and mortality. The objective is to develop a preliminary method for rapid antibiotic susceptibility testing (RAST) in enterobacteria with inducible chromosomal AmpC. METHODS: RAST was performed directly on spiked blood cultures of 49 enterobacteria with inducible chromosomal AmpC. Results were read at 4, 6 and 8h of incubation. Commercial broth microdilution was considered the reference method. Disks of 10 antibiotics were evaluated. RESULTS: The proportion of readable tests at 4h was 85%. All RAST could be read at 6 and 8h. For most antibiotics, the S or R result at 4, 6 and 8h was greater than 80% after tentative breakpoints were established and Area of Technical Uncertainty was defined. CONCLUSIONS: This preliminary method seems to be of practical use, although it should be extended to adjust the breakpoints and differentiate them by species.

Association of persistent positive blood cultures and infective endocarditis: A cohort study among patients with suspected infective endocarditis.

OBJECTIVES: To ascertain whether infective endocarditis (IE) was associated with persistent bacteraemia/candidaemia among patients with suspected IE. METHODS: This study included bacteraemic/candidaemic adult patients with echocardiography and follow-up blood cultures. Persistent bacteraemia/candidaemia was defined as continued positive blood cultures with the same microorganism for 48 h or more after antibiotic treatment initiation. Each case was classified for IE by the Endocarditis Team. RESULTS: Among 1962 episodes of suspected IE, IE (605; 31%) was the most prevalent infection type. Persistent bacteraemia/candidaemia was observed in 426 (22%) episodes. Persistent bacteraemia was more common among episodes with Staphylococcus aureus bacteraemia compared to episodes with positive blood cultures for other pathogens (32%, 298/933 vs 12%, 128/1029; P < 0.001). Multivariable analysis demonstrated that cardiac predisposing factors (aOR 1.84, 95% CI 1.31-2.60), community or non-nosocomial healthcare-associated (2.85, 2.10-3.88), bacteraemia by high-risk bacteria, such as S. aureus, streptococci, enterococci or HACEK (1.84, 1.31-2.60), two or more positive sets of index blood cultures (6.99, 4.60-10.63), persistent bacteraemia/candidaemia for 48 h from antimicrobial treatment initiation (1.43, 1.05-1.93), embolic events within 48h from antimicrobial treatment initiation (12.81, 9.43-17.41), and immunological phenomena (3.87, 1.09-1.78) were associated with infective endocarditis. CONCLUSIONS: IE was associated with persistent bacteraemia/candidaemia, along with other commonly associated factors.

Distinct patterns of vital sign and inflammatory marker responses in adults with suspected bloodstream infection.

OBJECTIVES: To identify patterns in inflammatory marker and vital sign responses in adult with suspected bloodstream infection (BSI) and define expected trends in normal recovery. METHODS: We included patients ≥16 y from Oxford University Hospitals with a blood culture taken between 1-January-2016 and 28-June-2021. We used linear and latent class mixed models to estimate trajectories in C-reactive protein (CRP), white blood count, heart rate, respiratory rate and temperature and identify CRP response subgroups. Centile charts for expected CRP responses were constructed via the lambda-mu-sigma method. RESULTS: In 88,348 suspected BSI episodes; 6908 (7.8%) were culture-positive with a probable pathogen, 4309 (4.9%) contained potential contaminants, and 77,131(87.3%) were culture-negative. CRP levels generally peaked 1-2 days after blood culture collection, with varying responses for different pathogens and infection sources (p < 0.0001). We identified five CRP trajectory subgroups: peak on day 1 (36,091; 46.3%) or 2 (4529; 5.8%), slow recovery (10,666; 13.7%), peak on day 6 (743; 1.0%), and low response (25,928; 33.3%). Centile reference charts tracking normal responses were constructed from those peaking on day 1/2. CONCLUSIONS: CRP and other infection response markers rise and recover differently depending on clinical syndrome and pathogen involved. However, centile reference charts, that account for these differences, can be used to track if patients are recovering line as expected and to help personalise infection.

Hemocultivos contaminados: bundle para lograr proporciones aceptables / Blood cultures contaminated: bundle to achieve acceptable proportions

Introducción. El problema de la contaminación de los hemocultivos es muy frecuente en establecimientos de atención hospitalaria, da lugar a la administración de antibióticos innecesarios y prolonga la hospitalización. Objetivo principal. Aplicar un bundle para reducir la proporción de contaminación de hemocultivos. Objetivo secundario. Realizar una encuesta anónima para detectar oportunidades de mejora en la técnica de extracción de hemocultivos. Metodología. Diseño del estudio: Estudio cuasi experimental que evaluó la proporción de contaminación de hemocultivos antes y después de implementar un bundle propio. Se determinó la proporción basal de contaminación de hemocultivos (ene-jul 2022), se realizó la intervención (agosto 2022) y se estableció la proporción de contaminación post intervención (sep.-abril 2023). Intervención: Se analizó la estructura, procedimiento y conocimiento del personal mediante una encuesta propia para detectar áreas de mejora. Se capacitó, a los técnicos de laboratorio, sobre el procedimiento de la toma de muestra mediante una simulación utilizando un brazo artificial. Se diseñó un bundle de seis medidas, se adaptó el procedimiento de toma de hemocultivo y se capacitó al personal. Análisis estadístico. Se analizó la proporción de hemocultivos contaminados entre los periodos pre y post utilizando Chi2 y la relación entre la proporción del periodo pre y post vs la literatura (3.00% contaminación aceptable) utilizando test Z para una proporción. Se consideró un p<0.05 como estadísticamente significativa. Se utilizo el software Stata 8. Resultados. Durante el estudio se analizaron un total de 3,965 hemocultivos. De estos, 1,978 corresponden al periodo pre-intervención y 1,987 corresponden al periodo post intervención. Durante la pre-intervención se detectaron 61 hemocultivos contaminados (3.08% vs 3.00% bibliografía, p:0.5866) mientras que en la etapa post intervención fue de 30 hemocultivos contaminados (1.51% vs 3.00% bibliografía, p:0.0000). La proporción de hemocultivos contaminados se redujo a la mitad, 3.08% vs 1.51%, p: 0.001. Se realizó una encuesta anónima pre y post intervención logrando mejoras en la técnica de toma de hemocultivos. Conclusión. La implementación del bundle propio para la extracción de hemocultivos, permitió reducir la proporción de contaminación a la mitad. El análisis de la encuesta nos permitió identificar oportunidades de mejora en la técnica de recolección de muestra de hemocultivos

Introduction: Contamination of blood cultures is very common in hospital care settings and results in the administration of unnecessary antibiotics and prolongs hospitalization. Main goal: Apply a bundle to reduce the rate of contamination of blood cultures. Secondary objective: Conduct an anonymous survey to detect opportunities for improvement in the blood culture extraction technique. Methodology: Study design: Quasi-experimental study that evaluated the proportion of blood culture contamination before and after implementing its own bundle. The baseline proportion of blood culture contamination was determined (Jan-July 2022), the intervention was performed (August 2022) and the post-intervention contamination proportion was established (September-April 2023). Intervention: The structure, procedure and knowledge of the staff was analyzed through an own survey to detect areas for improvement. Laboratory technicians were trained on the sample collection procedure through a simulation using an artificial arm. A bundle of six measures was designed: (hand hygiene with alcohol gel, use of common gloves and sterile gloves during extraction, antisepsis with alcoholic chlorhexidine gluconate, marking of the blood culture bottle up to the filling level, disinfection of the bottle cap). blood culture bottle with 70% alcohol, safety-lok kit with vacuum extraction system). The procedure was adapted and staff trained. Statistic analysis: The proportion of contaminated blood cultures between the pre and post periods was analyzed using Chi2 and the relationship between the proportion of the pre and post period vs the literature (3.00% acceptable contamination) using Z test for a proportion. P<0.05 was considered statistically significant. Stata 8 software was used.Results: A total of 3,965 blood cultures were analyzed during the study. Of these, 1,978 correspond to the pre-intervention period and 1,987 correspond to the post-intervention period. During the pre-intervention, 61 contaminated blood cultures were detected (3.08%) while in the post-intervention stage there were 30 contaminated blood cultures (1.51%). The proportion of contaminated blood cultures was reduced by half, 3.08% vs 1.51%, p: 0.001. An anonymous survey was carried out pre and post intervention, achieving improvements in the technique of taking blood cultures. Conclusion: The implementation of the own bundle for the extraction of blood cultures allowed the contamination rate to be reduced by ha

[Usefulness of the MPB-INFURG-SEMES model to predict bacteremia in the patient with solid tumor in the Emergency Department]. / Utilidad del modelo MPB-INFURG-SEMES para predecir bacteriemia en el paciente con neoplasia sólida en el Servicio de Urgencias.

OBJECTIVE: To analyse a new risk score to predict bacteremia (MPB-INFURG-SEMES) in the patients with solid tumor attender for infection in the emergency departments (ED). METHODS: Prospective, multicenter observational cohort study of blood cultures (BC) obtained from adult patients with solid neoplasia treated in 63 EDs for infection from November 1, 2019, to March 31, 2020. The predictive ability of the model was analyzed with the area under the Receiver Operating Characteristic curve (AUC-ROC). The prognostic performance for true bacteremia was calculated with the chosen cut-off for getting the sensitivity, specificity, positive predictive value and negative predictive value. RESULTS: A total of 857 blood samples wered cultured. True cases of bacteremia were confirmed in 196 (22.9%). The remaining 661 cultures (77.1%) wered negative. And, 42 (4.9%) were judged to be contaminated. The model's area under the receiver operating characteristic curve was 0.923 (95% CI,0.896-0.950). The prognostic performance with a model's cut-off value of ≥ 5 points achieved 95.74% (95% CI, 94,92-96.56) sensitivity, 76.06% (95% CI, 75.24-76.88) specificity, 53.42%(95% CI, 52.60-54.24) positive predictive value and 98.48% (95% CI, 97.66- 99.30) negative predictive value. CONCLUSIONS: The MPB-INFURG-SEMES score is useful for predicting bacteremia in the adults patients with solid tumor seen in the ED.

Isolation and identification of Wickerhamiella tropicalis from blood culture by MALDI-MS.

Wickerhamiella is a genus of budding yeast that is mainly isolated from environmental samples, and 40 species have been detected. The yeast isolated from human clinical samples usually only contain three species: W. infanticola, W. pararugosa and W. sorbophila. In this study, we isolated W. tropicalis from a blood sample of a six-year-old female with a history of B-cell precursor lymphoblastic leukemia in Japan in 2022. Though the strain was morphologically identified as Candida species by routine microbiological examinations, it was subsequently identified as W. tropicalis by sequencing the internal transcribed spacer (ITS) of ribosomal DNA (rDNA). The isolate had amino acid substitutions in ERG11 and FKS1 associated with azole and echinocandin resistance, respectively, in Candida species and showed intermediate-resistant to fluconazole and micafungin. The patient was successfully treated with micafungin. Furthermore, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) detected three novel peaks that are specific for W. tropicalis, indicating that MALDI-MS analysis is useful for rapid detection of Wickerhamiella species in routine microbiological examinations.

Rapid diagnostic of multidrug-resistant sepsis pathogens directly from blood culture bottles using MALDI-TOF and the EUCAST RAST.

In this study, rapid diagnostic of multidrug-resistant (MDR) sepsis pathogens, directly from positive blood culture (BC) bottles, was evaluated by combining MALDI-TOF and the EUCAST Rapid Antimicrobial Susceptibility Testing (RAST). Carbapenemase production in Escherichia coli and Klebsiella pneumoniae isolates was also evaluated by RAST. From 171 positive BC bottles analyzed, 79 (46 %) MDR species, including E. coli (4/34, 12 %), K. pneumoniae (33/48, 69 %), Pseudomonas aeruginosa (12/12, 100 %), Acinetobacter baumannii (15/15, 100 %), and Staphylococcus aureus (14/37, 38 %) displaying resistance to beta-lactams, fluoroquinolones, aminoglycosides, and/or trimethoprim/sulphamethoxazole, were identified. In this regard, turnaround time of direct MALDI-TOF identification and RAST was < 7 h, which was significantly (p< 0.05) lower than our routine method. Carbapenemase detection by RAST displayed 100% sensitivity and 88.7 % specificity at 8 h. This protocol could offer advantages for the treatment and clinical outcomes of septic patients, improving the rapid diagnostic of sepsis by MDR pathogens.

Rapid identification,fluconazole and micafungin susceptibility testing of Candida species from blood culture by a short incubation method.

This study aimed to develop and validate a rapid method for identification by MALDI-TOF system and determination of the susceptibility to Fluconazole and Micafungin by broth microdilution among Candidaspecies causing bloodstream infections. Subcultures from blood culture bottles were incubated for 5 hours (+/- 1h) and used to perform the tests, so that the turnaround time of rapid identification and susceptibility profile was about 5 and 24 hours, respectively. The rapid identification showed agreement of 92.05 %. Regarding the rapid broth microdilution for Fluconazole and Micafungin, the agreement was 97.06 % (p<0.001) and 100 % (p<0.001), and the Kappa coefficient was 0.91 (p<0.001) and 1.0 (p<0.001), respectively. To conclude, both rapid methods showed to be reproducible, inexpensive, easy to perform and time-saving. Thus, these methodologies could be useful to guide and adjust empirical antifungal therapy.

Analysis of antibiotic resistance of KPC-2-positive Klebsiella pneumoniae strains isolated from blood cultures in Van, Turkey.

OBJECTIVE: Recently, Klebsiella pneumoniae strains causing bacteremia and showing significant antibiotic resistance have raised serious health concerns. Especially carbapenem-resistant K. pneumoniae has spread worldwide and caused an increase in mortality rates. In this study, we aimed to present information about KPC-2 carriage and molecular characteristics of K. pneumoniae strains showing multiple antibiotic resistance among patients in our hospital. PATIENTS AND METHODS: Blood samples were collected from patients hospitalized in the intensive care units (ICU) of Van Regional Training and Research Hospital between 2020-2021. Culture, biochemical tests, antibiotic susceptibility tests, and molecular tests were performed to identify K. pneumoniae strains isolated from blood culture samples. RESULTS: Two hundred and sixteen K. pneumoniae were isolated from patients with positive blood cultures. Twenty-seven of these isolates showed multidrug resistance. Carbapenem, ß-lactam, and quinolone resistance were particularly high. On the contrary, almost all of these isolates were susceptible to Amikacin (AK), Gentamicin (CN), Colistin (CT) and Tigecycline (TGC). Molecular analysis revealed that all of these isolates were KPC-2-positive and ST11 variants. CONCLUSIONS: It was observed that KPC-2- positive K. pneumoniae strains with multi-drug resistance may pose a serious risk in patients hospitalized in ICU in our hospital. It was concluded that surveillance and personnel training regarding the hospital and community-acquired infections due to these isolates that show pandemic spread would be important.

Time to positivity of Klebsiella pneumoniae in blood cultures as prognostic marker in patients with intra-abdominal infection: A retrospective study.

Klebsiella pneumoniae is a common causative pathogen of intra-abdominal infection with concomitant bacteraemia, leading to a significant mortality risk. The time to positivity (TTP) of blood culture is postulated to be a prognostic factor in bacteraemia caused by other species. Therefore, this study aimed to investigate the prognostic value of TTP in these patients. The single-centred, retrospective, observational cohort study was conducted between 1 July 2016 and 30 June 2021. All adult emergency department patients with diagnosis of intra-abdominal infection and underwent blood culture collection which yield K. pneumoniae during this period were enrolled. A total of 196 patients were included in the study. The overall 30-day mortality rate was 12.2% (24/196), and the median TTP of the studied cohort was 12.3 h (10.5-15.8 h). TTP revealed a moderate 30-day mortality discriminative ability (area under the curve 0.73, p < 0.001). Compared with the late TTP group (>12 h, N = 109), patients in the early TTP (≤12 h, N = 87) group had a significantly higher risk of 30-day morality (21.8% vs. 4.6%, p < 0.01) and other adverse outcomes. Furthermore, TTP (odds ratio [OR] = 0.79, p = 0.02), Pitt bacteraemia score (OR = 1.30, p = 0.03), and implementation of source control (OR = 0.06, p < 0.01) were identified as independent factors related to 30-day mortality risk in patients with intra-abdominal infection and K. pneumoniae bacteraemia. Therefore, physicians can use TTP for prognosis stratification in these patients.

Streptococcus taonis sp. nov., a novel bacterial species isolated from a blood culture of a patient.

One Gram stain-positive, catalase-negative, α-hemolytic, chain-forming or paired cocci, designated ST22-14T, was isolated from a blood culture of a child with suspected infection. The results of 16S rRNA gene sequences analyses showed that the most closely related species to strain ST22-14T were "Streptococcus vulneris" DM3B3T (99.2%), Streptococcus mitis NCTC 12261T (99.0%), "Streptococcus gwangjuense" ChDC B345T, (99.0%), Streptococcus oralis subsp. dentisani 7747T (99.0%), Streptococcus downii CECT 9732T (99.0%), and Streptococcus infantis ATCC 700779T (98.9%). The genome of strain ST22-14T consists of 2,053,261 bp with a G + C content of 39.4%. Average nucleotide identity values between strain ST22-14T and Streptococcus mitis NCTC 12261T or other five species were from 82.2 to 88.0%. In silico DNA-DNA hybridization of ST22-14T showed an estimated DNA reassociation value of 34.6% with the closest species. The main cellular fatty acids of strain ST22-14T were 16:0, 18:0, 14:0, 18:1ω7c and 18:1ω6c. Based on these results, strain ST22-14T should be classified as a novel species of genus Streptococcus, for which the name Streptococcus taonis sp. nov. is proposed (type strain ST22-14T = NBRC 116002T = BCRC 81402T).

Analysis of Blood Cultures from Major Burns Patients in a Tertiary Care Burn Unit in Oman.

Objectives: This study aimed to review the blood stream infections of major burn patients in a tertiary care burn unit to determine the most prevalent infecting organisms in order to have a better empirical therapy protocol. Methods: This retrospective study analysed the blood stream infection of 155 major burn (>20% Total Body Surface Area [TBSA]) patients in Khoula Hospital, Muscat, Oman between January 2014 to December 2019. Results: The median age was 33 years and 57.42% of patients were male. The median TBSA was 38%, mortality was 25.16% and 50.9% of patients had positive blood cultures. The expired patients had higher TBSAs, Abbreviated Burns Severity Index scores and earlier first positive blood cultures. Candida was commonly grown in all the blood cultures, but the most prevalent organisms were Acinetobacter, Staphylococci, Klebsiella, Enterococcus and Pseudomonas. All Acinetobacter species are multidrug resistant. Of the 17 patients who had Kelbsiella grown in the blood culture, 8 grew multidrug-resistant Klebsiella. Only 4 patients' blood cultures grew methicillin-resistant Staphylococcus aureus. The number of blood culture samples taken ranged between 1-28 (median = 6). The first positive blood culture showed that Staphylococcus epidermidis and Acinetobacter were the most common infecting organisms. Conclusions: Multidrug-resistant Acinetobacter was the most predominant microorganism grown from the blood cultures of major burn patients in a tertiary care burn unit. Empirical therapy should include antibiotics that are effective against this organism to reduce the mortality.

Clinical utility of multiplex PCR in the detection of pathogens from sterile body fluids.

Rapid identification of pathogens in normally sterile body fluid (NSBF) is essential for appropriate patient management, specifically antimicrobial therapy. Limited sensitivity and increased time to detection of traditional culture prompted us to evaluate additional testing to contribute to the diagnosis of infection. The purpose of this study was to evaluate the GenMark Dx ePlex Blood Culture Identification (BCID) Panels on positive body fluids inoculated into blood culture bottles for the detection of microorganisms. A total of 88 positive body fluids from blood culture bottles were analyzed using a Gram-Positive, Gram-Negative, and/or Fungal pathogen BCID Panel based on the Gram stain result. Each result was compared to routine culture performed from the positive bottle. When using culture as a reference standard, we found the ePlex multiplex panel performed with a positive percent agreement of 96.5% and a negative percent agreement of 99.8%. The use of multiplex PCR may be a useful supplement to routine culture for NSBF in blood culture bottles. IMPORTANCE: The identification of pathogens in normally sterile body fluid (NSBF) is performed using routine culture, the current gold standard. Limitations of this method include sensitivity and increased turnaround times which could potentially delay vital patient care, especially antimicrobial therapy. Adaptations of NSBF in blood culture bottles prompted us to consider the utility of additional methods to bridge the gap in diagnostic challenges for these life-threatening infections. Multiplex molecular panels have been manufactured for use with multiple specimen types including blood, cerebral spinal fluid, stool, and respiratory. Therefore, the purpose of this study was to evaluate the off-label use of ePlex Blood Culture Identification Panels on positive body fluids grown in blood culture bottles for the detection of microorganisms for research purposes.