Routine Blood Cultures Research Articles

Case report: A case of blood culture-negative Bartonella quintana endocarditis: blood mNGS is an efficient method for early diagnosis

Bartonella quintana is one of the main causes of blood culture-negative endocarditis, and routine blood culture and serological methods are difficult to achieve early diagnosis. We report a case of blood culture-negative Bartonella quintana endocarditis from southwestern Chongqing. The patient was a 67-year-old male scavenger who presented with heart failure without fever as the main clinical manifestation upon admission. He stated having had contact with stray cats in the past 2 months. The combination of clinical symptoms, echocardiography, and blood mNGS testing confirmed the infection of Bartonella quintana.

A Comparative Study of Bacteriological Profile and Susceptibility Pattern by Rapid Direct Manual Identification and Antimicrobial Susceptibility Testing with Routine Blood Culture Method in Neonatal Septicemia.

A Comparative Study of Bacteriological Profile and Susceptibility Pattern by Rapid Direct Manual Identification and Antimicrobial Susceptibility Testing with Routine Blood Culture Method in Neonatal Septicemia.

Practices to prevent central line-associated bloodstream infection: A 2021 survey of infection preventionists in US hospitals.

To determine prevalence of technical and behavioral interventions aimed at preventing central line-associated bloodstream infection (CLABSI) following the COVID19 pandemic. Cross-sectional survey. US acute care hospitals. Infection preventionists at participating hospitals. Surveys were sent to infection preventionists from a national random sample of 881 US acute care hospitals. Questions covered use of technical interventions to prevent CLABSI (eg, alcohol-containing chlorhexidine gluconate [CHG] for skin antisepsis, use of coated catheters), socio-adaptive interventions (eg, feedback of CLABSI rates, use of appropriateness criteria), and leadership support for CLABSI prevention. Survey response rate was 47% (415/881). Technical interventions such as maximal sterile barriers (99%) or CHG-impregnated dressings (92%) were highly prevalent, but routine use of CHG bathing was less common (68% indicated regular use in intensive care unit [ICU] vs 18% in non-ICU settings). Although 97% of respondents indicated use of systems to monitor CLABSI, feedback to providers on CLABSI events was reported by 89%. Only 53% of respondents indicated regular use of tools to determine appropriateness of central venous catheters (CVC). Three-quarters of respondents indicated their hospital assessed CVC necessity daily, but only 23% reported strategies to reduce routine blood cultures. CLABSI prevention was extremely important to hospital leadership at 82% of responding hospitals. Most US hospitals continue to use evidence-based methods to prevent CLABSI as recommended by leading organizations. Opportunities to focus on socio-adaptive interventions such as feedback of infection rates, use of appropriateness criteria for CVC placement, and improving the "culture of pan-culturing" remain.

Post-COVID reactivation of latent Bartonella henselae infection: a case report and literature review

Cat scratch disease (CSD) is caused by Bartonella henselae (B. henselae) and presents as lymphadenopathy following close contact with cats. However, in context of the global COVID-19 pandemic, clinical manifestations of CSD may vary, posing new challenges for healthcare professionals. Here we describe a case of a 54-year-old male with painful left upper arm mass, which gradually resolved until he was infected with COVID-19. The mass then rapidly progressed before admission. Meanwhile, pulmonary symptoms including pleural effusion emerged simultaneously. The cause was undetermined with routine blood culture and pathological test until the next generation sequencing (NGS) confirmed the presence of B. henselae. We believe this case is the first to report localized aggravation of CSD after COVID-19 infection and hopefully, offers treatment experience for clinicians worldwide.

Development and proof-of-concept demonstration of a clinical metagenomics method for the rapid detection of bloodstream infection

BackgroundThe timely and accurate diagnosis of bloodstream infection (BSI) is critical for patient management. With longstanding challenges for routine blood culture, metagenomics is a promising approach to rapidly provide sequence-based detection and characterisation of bloodborne bacteria. Long-read sequencing technologies have successfully supported the use of clinical metagenomics for syndromes such as respiratory illness, and modified approaches may address two requisite factors for metagenomics to be used as a BSI diagnostic: depletion of the high level of host DNA to then detect the low abundance of microbes in blood.MethodsBlood samples from healthy donors were spiked with different concentrations of four prevalent causative species of BSI. All samples were then subjected to a modified saponin-based host DNA depletion protocol and optimised DNA extraction, whole genome amplification and debranching steps in preparation for sequencing, followed by bioinformatical analyses. Two related variants of the protocol are presented: 1mL of blood processed without bacterial enrichment, and 5mL of blood processed following a rapid bacterial enrichment protocol—SepsiPURE.ResultsAfter first identifying that a large proportion of host mitochondrial DNA remained, the host depletion process was optimised by increasing saponin concentration to 3% and scaling the reaction to allow more sample volume. Compared to non-depleted controls, the 3% saponin-based depletion protocol reduced the presence of host chromosomal and mitochondrial DNA < 106 and < 103 fold respectively. When the modified depletion method was further combined with a rapid bacterial enrichment method (SepsiPURE; with 5mL blood samples) the depletion of mitochondrial DNA improved by a further > 10X while also increasing detectable bacteria by > 10X. Parameters during DNA extraction, whole genome amplification and long-read sequencing were also adjusted, and subsequently amplicons were detected for each input bacterial species at each of the spiked concentrations, ranging from 50–100 colony forming units (CFU)/mL to 1–5 CFU/mL.ConclusionIn this proof-of-concept study, four prevalent BSI causative species were detected in under 12 h to species level (with antimicrobial resistance determinants) at concentrations relevant to clinical blood samples. The use of a rapid and precise metagenomic protocols has the potential to advance the diagnosis of BSI.

Microbiological characteristics of bloodstream infections in a reference hospital in northeastern Brazil.

Routine blood culture is used for the detection of bloodstream infections by aerobic and anaerobic bacteria and by common pathogenic yeasts. A retrospective study was conducted in a public hospital in Maceió-AL, by collecting data of all medical records with positive blood cultures. Out of the 2,107 blood cultures performed, 17% were positive with Staphylococcus coagulase negative (51.14%), followed by Staphylococcus aureus (11.21%) and Klebsiella pneumoniae (6.32%). Gram-positive bacteria predominated among positive blood cultures, highlighting the group of Staphylococcus coagulase-negative. While Gram-negative bacteria had a higher number of species among positive blood cultures.

Utility of Routine Blood Cultures for Upper Extremity Abscesses.

Patients with skin and soft tissue infections (SSTIs) are often admitted by the emergency department for intravenous antibiotic therapy and surgical drainage of abscesses if necessary. As part of the initial diagnostic workup, blood cultures are routinely drawn at our institution in patients with SSTIs. This study seeks to identify the utility of performing blood cultures in patients with upper extremity abscesses as it relates to the number of incision and drainage (I&D) procedures performed, patient readmission rates, and length of hospital stay. A retrospective chart review of 314 patients aged 18 to 89 years who underwent 1 or more I&D procedures of upper extremity abscesses were included in the study. Patient demographic data, comorbidities, laboratory values, wound and blood culture results, number of I&D procedures performed, length of stay, and readmission rates were evaluated. Increasing age and white blood count were associated with an increased number of I&Ds performed. Obtaining blood cultures, whether positive or negative, was associated with increased length of stay. There was no association between obtaining blood cultures and number of procedures performed on multivariable analysis. Positive blood cultures were associated with increased readmission rates. Routinely obtaining blood cultures in patients with upper extremity abscesses may not be beneficial. Obtaining blood cultures is not associated with an increased number of I&D procedures or readmission rates. Furthermore, obtaining blood cultures, regardless of positivity, is associated with increased lengths of hospital stay.

Evaluating blood culture collection practice in children hospitalized with acute illness at a tertiary hospital in Malawi.

Blood culture collection practice in low-resource settings where routine blood culture collection is available has not been previously described. We conducted a secondary descriptive analysis of children aged 2-23 months enrolled in the Malawi Childhood Acute Illness and Nutrition (CHAIN) study, stratified by whether an admission blood culture had been undertaken and by nutritional status. Chi-square test was used to compare the differences between groups. A total of 347 children were included, of whom 161 (46%) had a blood culture collected. Children who had a blood culture collected, compared to those who did not, were more likely to present with sepsis (43% vs. 20%, p < 0.001), gastroenteritis (43% vs. 26%, p < 0.001), fever (86% vs. 73%, p = 0.004), and with poor feeding/weight loss (30% vs. 18%, p = 0.008). In addition, hospital stay in those who had a blood culture was, on average, 2 days longer (p = 0.019). No difference in mortality was observed between those who did and did not have a blood culture obtained. Blood culture collection was more frequent in children with sepsis and gastroenteritis, but was not associated with mortality. In low-resource settings, developing criteria for blood culture based on risk factors rather than clinician judgement may better utilize the existing resources.

591. Positivity of Blood Culture on Automated Incubation System After 72 Hours and Clinical Significance from the Large United States Tertiary Care Center

Abstract Background Hospitals commonly use automated blood culture incubation systems that provide continuous monitoring for routine blood cultures. Detecting microorganisms early on can help optimize the utilization of antibiotics (ABx) and enable clinicians to de-escalate or discontinue ABx as necessary, making it an important aspect of antimicrobial stewardship. The BD BACTEC Fx, an automated system for blood culture incubation, holds blood culture bottles until growth is demonstrated or for five days unless an extended incubation period is requested by the infectious disease physician. We conducted a clinical evaluation of the significance of positive blood cultures after 72 hours of incubation time. Methods After receiving approval from the institutional review board (IRB), we obtained positive blood cultures from July 2017 to June 2020, which we then stratified based on positivity from day one to day five. To evaluate clinical significance, we retrospectively reviewed charts for positive blood culture results on day four and day five. An Infectious Diseases subspecialty team determined the clinical significance of each case. Results During the time frame of July 2017 to June 2020, there were 120,320 blood cultures taken. Out of these, 7,558 were determined to be positive, with 76 being positive on day 4 and 22 being positive on day 5. Upon further inspection of patients with positive blood cultures, it was discovered that repeat blood cultures continued to be positive due to either untreated sources or a high burden of bacteremia. It was also found that bottles containing fungus, particularly candida glabrata and candida albicans, tested positive on day 4 requiring treatment. Table 1 and Table 2 contain more detailed results. Blood cultures positive on day 4 Blood cultures positive on day 4 and clinical significance Blood cultures positive on day 5 Blood cultures positive on day 5 and clinical significance Conclusion In our study we have found that not all positive blood cultures at 72 hours translate into clinical scenarios requiring antibiotic therapy. In fact, prolonged antimicrobial treatment in such cases may lead to unnecessary exposure to antibiotics and increased risk of antibiotic resistance. It is important to consider the full context of clinical and laboratory findings when interpreting blood culture results, as well as any potential sources of contamination. Disclosures All Authors: No reported disclosures

Solitary splenic abscess caused by Salmonella typhi in an adult - A case report

A 71-year-old male was admitted to our hospital with complaints of fever, weakness and abdominal pain for 15 days. PET CT showed findings suggestive of a solitary splenic abscess. CT guided aspiration of the abscess was done and sent to lab for testing along with routine investigations and blood culture. Aspirate culture grew . The patient was treated with intravenous Cefepime followed by oral Cefixime along with supportive care. On follow up, complete resolution of the abscess was noted.

Utility of metagenomics next-generation sequencing in the diagnosis and treatment of severe infectious diseases in the intensive care unit.

Metagenomic next-generation sequencing (mNGS) is a new method that combines high-throughput sequencing and bioinformatics analysis. However, it has not become as popular due to the limited testing equipment and high costs and lack of family awareness with not much relevant intensive care unit (ICU) research data. To explore the clinical use and value of metagenomics next-generation sequencing (mNGS) in patients with sepsis in the ICU. We conducted a retrospective analysis of 102 patients with sepsis admitted to the ICU of Peking University International Hospital from January 2018 to January 2022. Based on whether mNGS was performed, the identified patients were divided into the observation group (n= 51) and the control group (n= 51), respectively. Routine laboratory tests, including routine blood test, C-reactive protein, procalcitonin, and culture of suspicious lesion specimens were performed in both groups within 2 hours after admission to the ICU, while mNGS tests were performed in the observation group. Patients in both groups were routinely given initial anti-infective, anti-shock, and organ support treatment. Antibiotic regimens were optimized in a timely manner according to the etiological results. Relevant clinical data were collected. The testing cycle of mNGS was shorter than that of the conventional culture (30.79 ± 4.01 h vs. 85.38 ± 9.94h, P< 0.001), while the positive rate of mNGS was higher than that of the conventional culture (82.35% vs. 45.1%, P< 0.05), with obvious superiority in the detection of viruses and fungus. There were significant differences in the optimal time of antibiotics (48 h vs.100 h) and length of ICU stay (11 d vs. 16 d) between the observation group and control group (P< 0.01) respectively, with no difference in 28-day mortality (33.3% vs. 41.2%, P> 0.05). mNGS is useful in the detection of sepsis-causing pathogens in the ICU with the advantages of short testing time and high positive rate. There was no difference in the 28-day outcome between the two groups, which may be related to other confounding factors such as small sample size. Additional studies with extended sample size are needed.

Highly multiplexed targeted sequencing strategy for infectious disease surveillance

BackgroundGlobal efforts to characterize diseases of poverty are hampered by lack of affordable and comprehensive detection platforms, resulting in suboptimal allocation of health care resources and inefficient disease control. Next generation sequencing (NGS) can provide accurate data and high throughput. However, shotgun and metagenome-based NGS approaches are limited by low concentrations of microbial DNA in clinical samples, requirements for tailored sample and library preparations plus extensive bioinformatics analysis. Here, we adapted molecular inversion probes (MIPs) as a cost-effective target enrichment approach to characterize microbial infections from blood samples using short-read sequencing. We designed a probe panel targeting 2 bacterial genera, 21 bacterial and 6 fungi species and 7 antimicrobial resistance markers (AMRs).ResultsOur approach proved to be highly specific to detect down to 1 in a 1000 pathogen DNA targets contained in host DNA. Additionally, we were able to accurately survey pathogens and AMRs in 20 out of 24 samples previously profiled with routine blood culture for sepsis.ConclusionsOverall, our targeted assay identifies microbial pathogens and AMRs with high specificity at high throughput, without the need for extensive sample preparation or bioinformatics analysis, simplifying its application for characterization and surveillance of infectious diseases in medium- to low- resource settings.

Neonatal sepsis: Challenges in data access, harmonisation and analysis to inform empirical antibiotic recommendations in South African neonatal units

&#x0D; &#x0D; &#x0D; Severe bacterial and fungal infections (sepsis) are a leading cause of mortality among neonates in sub-Saharan Africa. In African neonatal units, most sepsis episodes are hospital acquired, and caused by Gram-negative, antimicrobial-resistant pathogens. With rising rates of antimicrobial resistance (AMR) in neonatal sepsis pathogens, there is an increasing probability of inappropriate empirical antibiotic therapy (so called ‘bug-drug’ mismatch or discordant therapy), as confirmation of pathogen identity and antibiotic susceptibility pattern is either not available or delayed. The impact of AMR and discordant empirical treatment on neonatal sepsis outcomes in Africa is largely unknown owing to the lack of the infection surveillance data needed to derive empirical antibiotic treatment recommendations. The Antibiotics for Neonatal Sepsis in Africa (ANSA) study will address this data gap by pooling and harmonising available single-centre research datasets and/or routine laboratory blood culture data to determine which antibiotics should be used for empirical treatment of neonatal sepsis. However, many challenges hamper these datasets, including: variable laboratory methods for pathogen identification and susceptibility testing; difficulty in collating and harmonising data from multiple sites; problems of record linkage in the absence of a single patient identifier; lack of centralised health data capturing systems; and difficulty accessing data due to complex data sharing agreements and custodian permissions. To overcome some of these challenges, minimum guidelines for the collection of neonatal sepsis data need to be defined, to ensure consistency between datasets. Greater collection of, access to and sharing of neonatal sepsis data from institutional, provincial and national laboratories, as well as individual neonatal units, would enable the establishment of a neonatal surveillance network. Development of a sustainably-funded neonatal sepsis surveillance network is a national priority, to ensure effective, evidence-based antimicrobial treatment of hospitalised neonates in South Africa.&#x0D; &#x0D; &#x0D;

Neonatal sepsis: Challenges in data access, harmonisation and analysis to inform empirical antibiotic recommendations in South African neonatal units

&#x0D; &#x0D; &#x0D; Severe bacterial and fungal infections (sepsis) are a leading cause of mortality among neonates in sub-Saharan Africa. In African neonatal units, most sepsis episodes are hospital acquired, and caused by Gram-negative, antimicrobial-resistant pathogens. With rising rates of antimicrobial resistance (AMR) in neonatal sepsis pathogens, there is an increasing probability of inappropriate empirical antibiotic therapy (so called ‘bug-drug’ mismatch or discordant therapy), as confirmation of pathogen identity and antibiotic susceptibility pattern is either not available or delayed. The impact of AMR and discordant empirical treatment on neonatal sepsis outcomes in Africa is largely unknown owing to the lack of the infection surveillance data needed to derive empirical antibiotic treatment recommendations. The Antibiotics for Neonatal Sepsis in Africa (ANSA) study will address this data gap by pooling and harmonising available single-centre research datasets and/or routine laboratory blood culture data to determine which antibiotics should be used for empirical treatment of neonatal sepsis. However, many challenges hamper these datasets, including: variable laboratory methods for pathogen identification and susceptibility testing; difficulty in collating and harmonising data from multiple sites; problems of record linkage in the absence of a single patient identifier; lack of centralised health data capturing systems; and difficulty accessing data due to complex data sharing agreements and custodian permissions. To overcome some of these challenges, minimum guidelines for the collection of neonatal sepsis data need to be defined, to ensure consistency between datasets. Greater collection of, access to and sharing of neonatal sepsis data from institutional, provincial and national laboratories, as well as individual neonatal units, would enable the establishment of a neonatal surveillance network. Development of a sustainably-funded neonatal sepsis surveillance network is a national priority, to ensure effective, evidence-based antimicrobial treatment of hospitalised neonates in South Africa.&#x0D; &#x0D; &#x0D;

Bacteria in Extracorporeal Membrane Oxygenation Circuit Clots of a Patient With Persistent Bacteremia: A Case Report.

A neonate with pulmonary hypertension was supported with extracorporeal membrane oxygenation (ECMO). During ECMO support, the patient developed Enterococcus faecalis bacteremia, treated with targeted antibiotics. Despite the maximum dose of antibiotics, routine blood cultures remained positive throughout the ECMO treatment. A circuit change was performed due to buildup of thrombotic material and disseminated intravascular coagulation (DIC) inside the circuit. Thrombus formation was more extensive in the first than the second circuit. Gram-positive diplococci were present in all initial circuit clots and gram-positive masses surrounded by fibrin were found inside thrombi of the second circuit. Scanning electron microscopy (SEM) revealed a dense fibrin network with embedded red blood cells and bacteria in the first circuit. In the second circuit, SEM analysis revealed scattered micro thrombi. Polymerase chain reaction for identification of bacteria in the thrombus of the first circuit showed the same bacteria as found in blood cultures and did not achieve a sufficient signal in the second circuit. This case report shows that bacteria can nestle in thrombi of an ECMO circuit and that there is a rationale for a circuit change in a patient with persistent positive blood cultures and DIC.

Routine Blood Culture in the Emergency Department Worthy or Waste?

The authors declare no conflicts of interest.

Evaluation of a rapid combination disc test (RCDT) for direct phenotypic detection of extended-spectrum β-lactamase production in E. coli from positive blood culture bottles.

The spread of multi-resistant bacteria endangers the effectiveness of empirical antimicrobial treatment, particularly in Gram-negative bloodstream infections. Thus, rapid and reliable susceptibility testing has become a key challenge of modern microbiology. Here, we evaluated a combination disc test for rapid detection of ESBL production in Escherichia coli (rapid combination disc test, RCDT) directly from blood cultures. RCDT with discs containing cefotaxime and ceftazidime alone or in combination with clavulanic acid was validated using a cryo-collection of 96 third-generation cephalosporin-resistant (3GCR), whole-genome sequenced E. coli isolates spiked into blood culture bottles. All isolates were subjected to RCDT and rapid antibiotic susceptibility testing (RAST). Zone diameters were assessed after 4, 6 and 8 h of incubation. All isolates also underwent conventional combination disc testing. The real-life performance of RCDT was assessed by analysis of 306 blood cultures growing E. coli. Eighty of 90 (88.9%) ESBL-positive E. coli validation isolates were correctly identified by RCDT after 4 h of incubation. The detection rate increased to 100% after 6 and 8 h. RCDT was negative in six 3GCR E. coli isolates expressing class B or C β-lactamases. RCDT from routine blood cultures correctly classified all 56 ESBL producers and 245/250 ESBL-negative isolates after 4 h, resulting in 100% sensitivity and 98.8% specificity. RCDT is a reliable method for rapid ESBL detection in E. coli directly from positive blood cultures. RCDT might complement RAST to support antibiotic stewardship interventions and treatment decisions.

Can Blood Culture Contaminaton Cloud Fungal Positivity?

Objective: Blood culture (BC) is gold standard for diagnosis of fungemia. Contamination of BC is a major problem worldwide, since it may delay actual diagnosis. The aim of this study was to evaluate “contaminant” vials with a prolonged incubation (max 30 days) with antimicrobial supplemented media to observe any mycological growth. Materials and Methods: Routine BCs optained patients of XXX Hospital for a year period were included. Render BC System BC12-8 (Render Biotech Co. Ltd., Shenzhen, China) were used. Contaminated vials were reincubated and conventionally-inoculated weekly for four weeks total. In case of any growth, identifications were done by PhoenixTM 100 system (Becton Dickinson, MA, USA) with cornmeal tween 80 agar (RTA Laboratories, Kocaeli, Turkey). Antifungal susceptibility testing were applied with CLSI disk diffusion method. Results: 3.9% (235 sets and additional 138 vials) of total 6047 BC sets were contaminated. Only one vial from central venous catheter showed fungal growth within the first week of conventional inoculation (&gt;8th day of total incubation). The isolate was identified as Candida guilliermondii complex and susceptible to caspofungin. Latter set of this patient were positive in the 3rd day of incubation. Conclusion: International guidelines recommend

Diagnostic value of metagenomic next-generation sequencing in sepsis and bloodstream infection.

To evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) in sepsis and bloodstream infection (BSI). A retrospective analysis of patients diagnosed with sepsis and BSI at the First Affiliated Hospital of Zhengzhou University from January 2020 to February 2022 was conducted. All the patients underwent blood culture and were divided into mNGS group and non-mNGS group according to whether mNGS was performed or not. The mNGS group was further divided into early group (< 1 day), intermediate group (1-3 days), and late group (> 3 days) according to the time of mNGS inspection. In 194 patients with sepsis and BSI, the positive rate of mNGS for identifying pathogens was significantly higher than that of blood culture (77.7% vs. 47.9%), and the detection period was shorter (1.41 ± 1.01 days vs. 4.82 ± 0.73 days); the difference was statistically significant (p < 0.05). The 28-day mortality rate of the mNGS group (n = 112) was significantly lower than that of the non-mNGS group (n = 82) (47.32% vs. 62.20%, p = 0.043). The total hospitalization time for the mNGS group was longer than that for the non-mNGS group (18 (9, 33) days vs. 13 (6, 23) days, p = 0.005). There was no significant difference in the ICU hospitalization time, mechanical ventilation time, vasoactive drug use time, and 90-day mortality between the two groups (p > 0.05). Sub-group analysis of patients in the mNGS group showed that the total hospitalization time and the ICU hospitalization time in the late group were longer than those in the early group (30 (18, 43) days vs. 10 (6, 26) days, 17 (6, 31) days vs. 6 (2, 10) days), and the ICU hospitalization time in the intermediate group was longer than that in the early group (6 (3, 15) days vs. 6 (2, 10) days); the differences were statistically significant (p< 0.05). The 28-day mortality rate of the early group was higher than that of the late group (70.21% vs. 30.00%), and the difference was statistically significant (p = 0.001). mNGS has the advantages of a short detection period and a high positive rate in the diagnosis of pathogens causing BSI and, eventually, sepsis. Routine blood culture combined with mNGS can significantly reduce the mortality of septic patients with BSI. Early detection using mNGS can shorten the total hospitalization time and the ICU hospitalization time of patients with sepsis and BSI.

Can Prolonged Incubation of Negative Blood Cultures Show Fungal Positivity?

Objective: Standard duration of one set blood culture (BC) is a maximum of 5-7 days. The aim of this study was to evaluate “culture-negative” vials with a prolonged incubation (max 30 days) time and to observe any mycological growth. Materials and Methods: Routine BCs optained from adult patients of Balıkesir Atatürk City Hospital for a year period were included. Render BC System BC128 (Render Biotech Co. Ltd., Shenzhen, China) were used. Randomly selected vials were re-incubated for additional three weeks by conventional methods. In case of any growth, identifications were done by PhoenixTM 100 system (Becton Dickinson, MA, USA) with cornmeal tween 80 agar (RTA Laboratories, Kocaeli, Turkey). Antifungal susceptibility testing was applied with CLSI disk diffusion method. Results: A total of 6047 BC sets were obtained and randomly chosen 1040+122 negative sets (A and B groups, respectively) were included. In group A, none of them had fungal growth. In B (ongoing empiric antifungal ≤48h), only 2 sets showed significant fungal growth, which were observed in 7±2 days, and all strains were identified as C.glabrata complex and these patients were on empiric fluconazole (200 mg/day). One isolate was susceptible dose dependent, the other one was resistant for fluconazole. Latter sets of these fungemia patients showed positive signals in routine incubation period. Conclusion: Invasive fungal infections are increasingly encountered and isolation capacity and optimization of BCs are crucial. In this study, it was obviously observed that standard incubation period is satisfactory in order to detect almost all fungemia.