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Existing clinical methods for bacteria detection lack speed, sensitivity, and, importantly, point-of-care (PoC) applicability. Thus, finding ways to push the sensitivity of clinical PoC biosensing technologies is crucial. Here we report a portable PoC device based on lens-free interferometric microscopy (LIM). The device employs high performance nanoplasmonics and custom bioprinted microarrays and is capable of direct label-free bacteria ( E. coli) quantification. With only one-step sample handling we offer a sample-to-data turnaround time of 40 min. Our technology features detection sensitivity of a single bacterial cell both in buffer and in diluted blood plasma and is intrinsically limited by the number of cells present in the detection volume. When employed in a hospital setting, the device has enabled accurate categorization of sepsis patients (infectious SIRS) from control groups (healthy individuals and noninfectious SIRS patients) without false positives/negatives. User-friendly on-site bacterial clinical diagnosis can thus become a reality.

Background: Culture-based methods (culture broth bottles or lysis methods) have been the standard for detecting bacteremia. More recently, quantitative polymerase chain reaction (qPCR) was proposed as a more sensitive and specific test although none of them has been validated for the identification of periodontal pathogens (fastidious growing bacteria) in blood samples., Objective: To compare the ability to detect and quantify Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Streptococcus oralis (alone or in combination) in blood samples with three culture techniques [direct anaerobic culturing (DAC), haemo-culture (BACTEC), and lysis-centrifugation (LC)] and a non-culture dependent approach (qPCR) in an in vitro study., Material and Methods: Blood samples from 12 periodontally healthy volunteers were contaminated with three concentrations [10 4 ,10 2 and 10 1 colony forming units (CFU)/mL] of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination. Samples were analysed by DAC, BACTEC, LC and qPCR. Sensitivity, specificity, predictive values, kappa index and Lińs correlation coefficients were calculated., Results: DAC, LC and qPCR were able to detect the three target species at all concentrations. An excellent concordance (correlation coefficient r: 0.92-1) was observed between DAC and the reference standard (sensitivity raging 93.33-100% and specificity 88.89-100%) values. BACTEC was not able to identify P. gingivalis in any of the performed experiments. qPCR provided false negative results for S.oralis., Conclusions: DAC showed the best results for the proper identification and quantification of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination, in blood samples., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Aims: Comparison of conventional blood culture with BACTEC 9050 for rate and time to detection of microorganisms., Settings and Design: A prospective study was carried out in a multispecialty tertiary care teaching hospital., Subjects and Methods: A total of 835 paired specimens (797 blood and 38 nonblood specimens) were collected and processed according to standard microbiological procedures by both conventional method as well as by BACTEC 9050 automated culture system. Clinical details of patients were recorded. Data were analyzed for time to detection and isolation rate by the two systems and compared., Results: Overall culture positivity for BACTEC 9050 and the conventional system was 32% and 19.88%, respectively. Eighty-five demonstrated concordant growth, 136 specimens were culture positive by BACTEC only, and 38 specimens were culture positive by conventional only. Twelve contaminants in BACTEC and nine contaminants in conventional system were detected. Using BACTEC 9050, higher isolation was observed for Acinetobacter spp., coagulase negative Staphylococcus spp., Streptococcus spp., and Candida spp. A total of 410 patients were on antimicrobial treatment and culture positivity was significantly higher with BACTEC 9050 (P < 0.0001). There was a significant difference in the mean time to detection with BACTEC 9050 recovering 86.8% of isolates within 48 h (P < 0.0001)., Conclusions: Although BACTEC 9050 demonstrated a significantly higher recovery of microorganisms from blood, an appropriately performed conventional blood culture can facilitate the choice of therapy.

A prospective observational study was conducted in two hospitals of Buenos Aires city (Argentina); 191 clinically significant monomicrobial gram-negative bloodstream infections were included in the study, which combined the Bact-Alert System Blood culture machine and the Vitek 2C System. Organism identification and susceptibility results directly from blood culture bottles were compared with those obtained from cards inoculated with a standardized bacterial suspension obtained following subculture on agar. By comparing the results obtained from pure cultures with those by the Vitek 2C System as reference method, the agreement between the reference method and the direct identification from positive blood cultures was 99 %. By antimicrobial susceptibility testing, the overall categorical accuracy was 99 % (0.22 %, very major errors, 0.17 %, major errors and 0.61 %, minor errors). One hundred and eight (56,8 %) bloodstream infections were treated empirically with adequate antibiotics. After the results obtained directly from the bottles were reported, antimicrobial therapy was changed in 116 (60.7 %) of the episodes.

Background: To evaluate the VersaTREK (TREK Diagnostic Systems, Cleveland, Ohio) blood culture system against the Bactec9240 (BD Microbiology, Cockeysville, MD), for the recovery of bloodstream pathogens., Methods: Venous blood from patients with suspected bacterial sepsis was evenly distributed into bottles of each system. Positive signals were recorded and bottles processed onto standard media for organism recovery. False positive signals were regarded if no organisms were seen on Gram stain and no growth was observed., Results: 177 bottles were available for analysis; the Bactec9240 system yielded 43 positive, 134 negative results and no false positive signals. The VersaTREK system had 58 positive signals with 14 being false positives., Conclusions: In our setting with high background burden of immuno-compromised patients, the VersaTREK system compared favourably with the Bactec9240 in recovering blood stream aerobic and facultative anaerobic pathogens from patients with suspected bacterial sepsis. A concern is the high false positivity rate. Due to its versatility to accommodate small and large workloads as well as using smaller volumes of blood, this system may establish itself as a useful alternative for the recovery of bloodstream pathogens.

The publications in 1990s have indicated decreased recovery rates of obligate anaerobes from blood cultures and have questioned the need for routine anaerobic blood culture bottles. In this study, we compared positivities of the paired aerobic and anaerobic bottles and rapidity to detect positive cultures by two automated blood culture systems, BACTEC 9120 and BacT/ALERT 3D. Of 401 positive readings by BACTEC 9120, 338(84.3%) aerobic bottles became to be positive, and anaerobic bottles were 318(79.3%). Also, of 437 positive readings by BacT/ALERT 3D, positivities were 90.8% and 67.3% by aerobic and anaerobic bottles, respectively. These results indicated 5.0% and 23.7% more organisms were recovered in aerobic bottles than in anaerobic bottles, including more staphylococci, gram-positive rods, glucose-nonfermentative gram-negative rods and yeasts. Only 4 (0.14%) of 2,799 BACTEC 9120 anaerobic bottles and 2 (0.06%) of 3,428 BacT/ALERT 3D anaerobic bottles recovered obligate anaerobes. We compared time to detect positive cultures during incubation cycle by both aerobic and anaerobic bottles. Aerobic bottles in BACTEC 9120 read more positive cultures >2 hours earlier than anaerobic bottles, whereas BacT/ALERT 3D could not demonstrate a statistical significance in rapid reading of positive cultures. These results support that recovery rates of obligate anaerobes markedly decreased and that the routine use of anaerobic blood culture bottles is not legitimate at this time. In place of anaerobes, it is an urgent and important issue how to recover fungi correctly and rapidly from blood cultures.

We hereby report two instances of dimorphic fungus cultivation in BacT/ALERT-based bacteriologic media, with the first such characterization of Blastomyces dermatitidis. From a patient with disseminated coccidioidomycosis, routine blood cultures incubated on the MB/BacT 3D Microbial Detection System generated a positive signal following 75 h of incubation. B. dermatitidis was isolated from a patient hospitalized with a four-week course of respiratory illness. Organism detection from respiratory specimens via the MB/BacT 3D Mycobacteria Detection System occurred 5 days sooner than the routine fungus culture. Etiologic agents of endemic mycoses may be isolated in bacteriologic media employed by continuous monitoring instrumentation.

Context: The optimal use of dedicated fungal and mycobacterial blood culture bottles, such as the BACTEC Myco/F Lytic bottle, has not been well defined in clinical practice., Objectives: To compare the performance of Myco/F Lytic and standard blood culture in clinical practice in an urban tertiary care hospital setting and to implement a strategy for optimal use of Myco/F Lytic culture., Data Sources: Retrospective review of laboratory records., Results: Myco/F Lytic culture did not increase detection of yeasts. Nor did it decrease time to detection except for Candida glabrata, where mean time to positivity dropped from 2.6 +/- 1.1 days in standard to 1.8 +/- 0.8 days in Myco/F Lytic culture. Therefore, an algorithm was developed in which Myco/F Lytic culture was reserved primarily for detection of mycobacteria in patients with severely depressed CD4 counts. Implementation of this algorithm led to a sustained 3-fold reduction in Myco/F Lytic blood culture usage., Conclusions: Retrospective analysis suggests substantial clinical equivalence of standard blood and Myco/F Lytic culture for detection of yeast. A multifaceted educational approach based on this data led to a sustained change in physician ordering practices and more cost-effective use of resources.

Blood culture is the cornerstone of an established aetiological diagnosis of septicaemia. The automated blood culture systems used for this purpose have changed little in the last decade, and the clinical value of results depends on a variety of factors, including pre- and post-analytical variables. Growing scepticism over the value of blood culture results and pressure for the introduction of molecular detection systems have prompted a critical path analysis of pre-, peri- and post-analytical stages in the generation of positive blood culture results. The impact of a positive blood culture was studied in a teaching hospital for 12 months before and 12 months after the introduction of a microbiologist's blood culture round. Active culture reporting via a blood culture ward round was supported by a personal data assistant database of contemporaneous laboratory and clinical data. Hospital occupancy and death register records were subsequently obtained through the State Government data linkage project. There was no evidence that faster laboratory generation of positive blood culture results, faster reporting of results or direct clinical interaction with the patient's primary medical team reduced the risk of death in hospital. However, there was a threefold increase in the rate of death in hospital following a 1 day delay in collection of blood cultures after hospital admission (P=0.0010). The overall duration of hospital stay for patients with a positive blood culture fell by 2.5 days compared with the previous 12 month period (P=0.0003). The interval between the initial positive culture result and patient discharge fell by 2 days (P=0.0010). This difference was attributed to shorter overall admissions and shorter intervals between positive cultures containing Gram-positive cocci and subsequent patient discharge (P=0.0018). An increased mortality rate from community-acquired bacteraemic infections was associated with delayed culture collection, but not with a prolonged laboratory processing interval. Thus, the speed of conventional blood culture analysis and the form of clinical reporting have little direct effect on the clinical outcome of bacteraemia, but may contribute to a reduction in the length of hospital admission. Introduction of molecular identification tests, such as multiplex PCR methods, at the Gram-stain stage of blood culture is unlikely to affect the rate of death in hospital, but may reduce the length of hospital admission.

Blood culture bottles with antimicrobial removal systems are recommended for patients who develop fever while on antibiotics. This study compared the ability of Becton Dickinson (Sparks, MD) BACTEC PLUS bottles and bioMerieux (Durham, NC) BacT/Alert FA bottles to effectively remove vancomycin, cefoxitin, ceftriaxone, cefepime, piperacillin-tazobactam, ampicillin, oxacillin, gentamicin, and a combination of gentamicin/penicillin, thus allowing bacterial pathogens to grow. Each bottle was spiked with 10 ml of human blood, antibiotic, and strains of organisms susceptible to the antibiotic evaluated. The organisms used were type strains and clinical isolates of Staphylococcus aureus (methicillin susceptible and resistant), Streptococcus pneumoniae, a viridans streptococcus, Enterococcus faecalis, Enterococcus faecium, Streptococcus agalactiae, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Testing was completed in triplicate, using 10 to 100 CFU/ml of organisms with various concentrations of each antibiotic. Two rounds of testing were completed per antibiotic/organism combination. Bottles were mixed and loaded onto their respective instruments as per the manufacturer's instructions. Antimicrobial removal was evaluated on the basis of time to detection of organism growth, for up to 5 days of incubation. Overall, the BacT/Alert FA system recovered 25.1% of strains from test bottles and 96.9% of strains from growth control bottles (no antibiotic added), and the BACTEC PLUS system recovered 95.1% of strains from test bottles and 100% of strains from growth control bottles. Both systems performed well in the detection of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa in the presence of gentamicin. In the presence of ceftriaxone, neither system was able to recover Streptococcus pneumoniae. The ability to remove vancomycin and cefoxitin was also determined by measuring antibiotic levels remaining in bottles after 1 h of incubation. The results demonstrated remaining levels of 72 to 90% of vancomycin and 71 to 72% of cefoxitin in the BacT/Alert system. For the BACTEC system, remaining levels were 0 to 30% of vancomycin and 0% of cefoxitin. Under these simulated conditions, the BACTEC PLUS system was superior to the BacT/Alert FA system in recovering gram-positive and gram-negative bacterial pathogens in the presence of beta-lactam antibiotics, gentamicin/penicillin, and vancomycin.

We compared the performance of the BacT/ALERT automated blood culture system with real-time, quantitative volatile organic compound (VOC) detection by selected ion flow tube-mass spectrometry (SIFT-MS). Blood samples from healthy donors were artificially infected with 5 or 100 CFU of organisms commonly causing bacteremia. Positive results by SIFT-MS analysis of headspace gases were recorded for 53/60 (88.3%) bottles at 8h, and 58/60 (96.6%) bottles at 24 h. We conclude that SIFT-MS is a sensitive method for the detection of microbial VOCs. Furthermore, profiles of the VOCs detected may allow simultaneous identification of infecting organisms.

This study explored the possibility of combining direct inoculation of tube coagulase and DNase tests, and the VITEK2 system, from BACTEC blood culture bottles in order to achieve rapid identification and susceptibility testing of Staphylococcus aureus. All isolates were identified correctly as S. aureus or coagulase-negative staphylococci (CNS). Antimicrobial susceptibility testing with the VITEK2 system gave 99.6% correct category agreement, with 0.1% very major errors and 0.3% minor errors among S. aureus isolates, and 97.4% correct category agreement, with 0.9% very major errors and 1.7% minor errors among CNS isolates. The results suggested that direct identification and susceptibility testing is sufficiently accurate for immediate reporting.

Rapid detection of micro-organisms from blood is one of the most critical functions of a diagnostic microbiology laboratory. Automated blood-culture systems reduce the time needed to detect positive cultures, and reduce specimen handling. The false-positive rate of such systems is 1-10%. In this study, the presence of pathogens in 'false-positive' bottles obtained from BACTEC 9050 (Becton Dickinson) and BacT/Alert (Biomérieux) systems was investigated by eubacterial and fungal PCR. A total of 169 subculture-negative aerobic blood-culture bottles (104 BacT/Alert and 65 BACTEC) were evaluated. Both fungal and eubacterial PCRs were negative for all BACTEC bottles. Fungal PCR was also negative for the BacT/Alert system, but 10 bottles (9.6%) gave positive results by eubacterial PCR. Sequence analysis of the positive PCR amplicons indicated the presence of the following bacteria (number of isolates in parentheses): Pasteurella multocida (1), Staphylococcus epidermidis (2), Staphylococcus hominis (1), Micrococcus sp. (1), Streptococcus pneumoniae (1), Corynebacterium spp. (2), Brachibacterium sp. (1) and Arthrobacter/Rothia sp. (1). Antibiotic usage by the patients may be responsible for the inability of the laboratory to grow these bacteria on subcultures. For patients with more than one false-positive bottle, molecular methods can be used to evaluate the microbial DNA in these bottles. False positives from the BACTEC system may be due to elevated patient leukocyte counts or the high sensitivity of the system to background increases in CO(2) concentration.

To validate performance, we compared the new plastic BacT/ALERT (bioMérieux, Durham, NC) SN bottle to the current glass SN bottle with samples of blood obtained for culture from adults and found them comparable for both recovery and speed of detection of microorganisms. We conclude that the safety advantage of plastic bottles can be achieved without compromising performance.

Septicaemia is a major contributor of mortality. Blood culture is the essential investigation for the management of sepsis. Due to lack of resources blood culture is an irregularly used investigation in India. A three-tier level of development is being proposed to develop the blood culture based national programme for early detection of sepsis. The plan envisages the establishment of manual blood culture based elementary system in the health centre and district hospital level (Level 1), direct Gram stain and direct antibiotic sensitivity testing from the "positive" blood culture broths at the medical college hospital level (Level 2) and development of automated methods, enhancement of quality control and safety measures, clinical liaison and re-orientation of microbiology training at the tertiary care centre level (Level 3).

The purpose of this study is to deal with the problem of anaerobic cultivation of clinical specimens and consider the possibility of using semi-automated blood culture instruments. The Bactec Lytic, BacT/Alert FAN Anaerobic and BacT/Alert FN Anaerobic bottles were inoculated with Bacteriodes fragilis, Clostridium perfringens, Peptostreptococcus anaerobius and Feingoldia magna strains. The times to detection (TTD) for positive bottles were evaluated with reference to the number of inoculated bacteria. Inoculation with the same suspension of equal bacterial strain resulted in shorter TTD values for all the Bactec Lytic bottles as compared with the BacT/Alert FAN Anaerobic or BacT/Alert FN Anaerobic bottles, respectively. Statistically significant differences were recorded (p=0,05) by Bacteroides fragilis and Clostridium perfringens species. Feingoldia magna was submitted to culture in the Bactec Lytic alone. The anaerobic blood culture bottles are deemed acceptable for application in the culture of anaerobc isolates drawn from other samples except for blood (namely when prompt delivery of the sample to the laboratory is impossible and the sample can suffer damage), however, only a limited range of microorganisms can be taken into account for the detection purposes. The Bactec Lytic bottles are more appropriately designed for the detection of anaerobic bacterial species compared to the BacT/Alert FAN Anaerobic and BacT/Alert FN Anaerobic bottles. They achieve faster growth parametres and provide more successful readings of anaerobic bacteria culture and detection.

Bacterial contamination of blood components represents today the highest infectious risk in blood transfusion, the risk is particularly high when it affects platelet concentrates. The prevention methods developed over the past ten years (donor selection, phlebotomy site preparation, first 30 ml diversion, systematic leuko-reduction...) which aimed at limiting the introduction of bacteria in donations and bacterial proliferation, has reduced the risk of transfusion reaction due to the bacterial contamination. Improving strategies for reducing the risks of bacterial contamination is one of the priorities of the French National Blood Transfusion Service (l'Etablissement français du sang - EFS). It is essential to improve existent prevention methods and develop the implication of all the actors (from donation to transfusion) involved in the prevention of this risk. Bacterial detection or pathogens inactivation are now available and are able to reduce (for detection) or prevent (for inactivation) the occurrence of reaction due to bacterial contamination of PC. Up to now, the choice is in favour of bacterial detection. Three methods (BacT/Alert, BioMerieux; eBDS, Pall; ScanSystem, Hemosystem) of detection of bacterial contamination in PC can be generalised now. Adaptations, need for their implementation are acceptable, especially concerning PC availability.

The plastic pediatric BacT/ALERT (bioMérieux, Durham, N.C.) PF (PPF) is a new nonvented aerobic culture medium in a clear plastic bottle designed to prevent breakage. We compared the performance of the new PPF bottle to that of the present glass BacT/ALERT PF bottle for the recovery of microorganisms as well as for the time to detection of growth in samples of blood obtained for culture from children. We found that the PPF and PF bottles were comparable for recovery of microorganisms and that the safety advantage of plastic bottles can be achieved without compromising performance.

The effects of volume of blood, number of consecutive cultures, and incubation time on pathogen recovery were evaluated for 37,568 blood cultures tested with the automated BACTEC 9240 instrument (Becton Dickinson Diagnostic Instrument Systems) at a tertiary care center over the period of 12 June 1996 through 12 October 1997. When the results for this study were compared with previous data published for manual broth-based blood culture systems and patient samples obtained in the 1970s and 1980s, the following were found: (1) the percentage increase in pathogen recovery per milliliter of blood is less, (2) more consecutive blood culture sets over a 24-h period are required to detect bloodstream pathogens, and (3) a shorter duration of incubation is required to diagnose bloodstream infections. Guidelines developed in the 1970s and 1980s for processing and culturing blood may require revision.

The present study describes the use of the automated BACTEC 9240 blood culture system, the Serum Separator Tube (SST), and the BD PHOENIX Automated Microbiology System in combination for the direct identification and antimicrobial susceptibility testing (AST) of gram-negative rods (GNRs) from positive blood cultures (BCs) without subculture. The study was conducted in three phases: (i) the recovery yield of Escherichia coli ATCC 25922 was determined with the SST between 0 and 8 h after spiked BC bottles turned positive; (ii) the identifications and susceptibility testing results obtained with the PHOENIX system for nine American Type Culture Collection strains of GNRs processed by the SST procedure and for colonies from agar medium were compared; and (iii) the procedure with the BACTEC system, SSTs, and the PHOENIX system was applied to positive cultures of blood from 309 patients during a 3-month period. The SST procedure with E. coli yielded sufficient numbers of cells to perform direct inoculation at any time between 0 and 8 h after a BC bottle turned positive. By using the identities obtained from pure cultures with the PHOENIX system and other biochemical identification systems as reference methods, the agreement between the reference methods and the PHOENIX system tested directly by using cultures of blood from patients was 92.9%. The 7.1% discrepant results were due to 6.5% incorrect identifications with the PHOENIX system with BC samples and 0.6% incorrect identifications with the PHOENIX system with samples from agar cultures. By AST the overall categorical accuracy was 99.0%, with 0.1% very major errors, 0.1% major errors, and 0.8% minor errors. In conclusion, use of the combination of the BACTEC system, SSTs, and the PHOENIX system has the potential to allow the agar isolation step to be skipped and the procedures for rapid direct identification and susceptibility testing of GNRs from positive BCs to be improved both in hospital-based and in central non-hospital-based laboratories.

Incubation periods of aerobic (AE) and anaerobic (AN) blood-culture bottles with the BacT/Alert system were assessed in our laboratory. We reviewed the results of 6229 blood-culture sets collected at Kyoto University Hospital from January 1999 to December 2000. Of these sets, 731 (11.7%) were positive for bacteria or yeast. Excluding 87 sets with growth evidence on arrival, of the 644 positive blood-culture sets from 341 patients, a total of 691 organisms were isolated. Of the 691 organisms, 413 (59.8%) were recovered from both bottles, 206 (29.8%) were recovered only from the AE bottle, and 72 (10.4%) were recovered only from the AN bottle. The AE bottle was significantly superior to the AN bottle in terms of both recovery rate and detection time for overall organisms, but there was no significant difference in detection time for facultative anaerobic bacteria between the two bottles. Of the 691 organisms, 530 (76.7%) were classified as usual pathogens. Of the 530 usual pathogens, 501 (94.5%) were recovered in at least one bottle of each set within the first 3 days, and 523 (98.7%) within the first 5 days of incubation. Twenty-nine organisms initially isolated on day 4 or later were recovered from 19 patients. Of these, chart reviews indicated that 21 organisms recovered from 11 patients were considered clinically significant bacteria, and the reviews also revealed that no patient had a treatment plan altered based on the results of positive blood culture. Seven organisms initially isolated on day 6 or later were recovered from 7 patients. Chart reviews revealed that 5 of these organisms from 5 patients were considered to be clinically significant. In conclusion, if the incubation period had been less than 3 days, 11 patients with clinically significant bacteremia or fungemia, (3.2% of all patients with bacteremia or fungemia) would have been undiagnosed. Similarly, with an incubation period of 5 days, 5 such patients (1.5%) would have been undiagnosed.

The BACTEC MYCO/F Lytic blood culture bottle (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) is designed to optimize the recovery of fungi and mycobacteria; however, this bottle also supports the growth of most aerobic bacteria. We compared the MYCO/F Lytic bottle with two other BACTEC bottles and the Isolator system for the recovery of bacteria as well as fungi and mycobacteria from blood. A total of 6,108 blood culture sets were inoculated with blood obtained from adult patients. Twenty-five to 28 ml of blood collected by a phlebotomy team for each blood culture set was randomly distributed into each of four blood culture receptacles: the Isolator tube (Wampole Laboratories, Cranbury, N.J.) and three BACTEC bottles: the MYCO/F Lytic bottle, the BACTEC Plus Aerobic/F bottle, and the BACTEC Anaerobic Lytic/10 bottle. The sediment from the Isolator tube was inoculated onto chocolate agar (CA), brain heart infusion agar (BHI), and Sabouraud dextrose agar (SDA) and into a BACTEC 13A bottle. Incubation durations were as follows: MYCO/F Lytic bottle, 42 days; Plus Aerobic/F bottle, 5 days; Anaerobic Lytic/10 bottle, 5 days; sediment from Isolator tube on CA, 3 days; sediment from Isolator tube on BHI, 30 days; sediment from Isolator tube on SDA, 30 days; and sediment from Isolator tube in a BACTEC 13A bottle, 42 days. Two isolates of Histoplasma capsulatum were recovered from the Isolator tube only. Three isolates of Mycobacterium tuberculosis complex were recovered: two isolates from the MYCO/F Lytic bottle only and one isolate from the Isolator tube (whose sediment was inoculated into the BACTEC 13A bottle) only. Two isolates of Cryptococcus neoformans were recovered: one from the MYCO/F Lytic bottle only and the other from the MYCO/F Lytic bottle and the Isolator tube (whose sediment was inoculated into the BACTEC 13A bottle). For potential pathogens overall, there was a statistical difference in recovery that favored the Isolator system over the MYCO/F Lytic bottle (P = 0.0015), including statistically significant differences for Staphylococcus aureus (P = 0.0001) and Streptococcus pneumoniae (P = 0.0313). However, there was no statistically significant difference between the two blood culture systems when detection of bloodstream infection was considered. The time to detection for all potential pathogens combined was less for the MYCO/F Lytic bottle than for the Isolator system (P = 0.0004). Overall, the potential pathogen recovery was greater for the BACTEC Plus Aerobic/F bottle than for either the Isolator system (P = 0.0003) or the MYCO/F Lytic bottle (P = 0.0001). However, the BACTEC Plus Aerobic/F bottle did not recover M. tuberculosis, H. capsulatum, or C. neoformans isolates. The combination of the Isolator system and MYCO/F Lytic bottle may be useful as a selective blood culture method to optimize the recovery of fungi and mycobacteria from blood. Compared with the manual Isolator system, the MYCO/F Lytic system has the advantage of less preanalytic processing and continuous automated monitoring of bottles for growth by the BACTEC 9240 instrument.

The current BacT/Alert standard aerobic (VA) blood culture bottle was redesigned and designated a nonvented aerobic (NVA) culture bottle; this bottle does not require venting. A total of 3,873 sets of blood samples for culture were obtained from adult patients with suspected bacteremia or fungemia. The NVA bottle showed performance equivalent to that of the VA bottle for recovery and speed of detection of microorganisms from blood without the need for venting the bottle.

Background: Neonatal sepsis is a low incidence, high-risk disease with many sepsis work-ups performed to detect a single case. Seventy-two hours of antibiotic therapy have been traditionally recommended pending negative culture results. Improved culture media and new technology integrated into blood culture systems could shorten incubation time required to detect positive culture results. This would then change the length of antibiotic therapy in the management of the newborn infant with suspected sepsis. In addition, previous data supporting the 72-hour recommendation were retrospectively acquired, utilized nonautomated systems, and reported in an era with a different population of microorganisms cultured in special care nurseries., Objective: Evaluate the time of incubation to detect positive blood cultures from newborn infants with suspected sepsis using a computer-assisted, automated blood culture system, ESP (Trek Diagnostic Systems, Inc, Westlake, OH)., Design: Prospective, observational study., Patients and Setting: All positive blood culture results that were obtained from term and preterm newborn infants born from November 1993 through June 1997 at a publicly funded hospital with over 6000 live births per year., Methods: As positive blood culture results were identified, data were prospectively obtained from the patient's medical record. The computer algorithm in the automated blood culture system determined the time to positivity. Time to positivity was determined for blood cultures obtained before the initiation antimicrobial therapy and compared with those cultures obtained after beginning therapy. Time to positivity was also evaluated for clinically important Gram-positive and Gram-negative bacteria and yeast., Results: Four hundred fifty-five positive blood culture results were obtained from 222 patients. Gram-positive organisms accounted for 80% (366/455) of the positive culture results, Gram-negative organisms accounted for 11% (48/455), and yeast for 9% (41/455). Virtually all cultures growing clinically significant Gram-positive and Gram-negative organisms were positive by 24 to 36 hours of incubation. Cultures growing Staphylococcus epidermidis were virtually all positive after 36 to 48 hours of incubation. Of cultures growing yeast, 88% (36/41) were positive by 48 hours of incubation. There was no difference in time to positivity in pretherapy or posttherapy obtained positive blood cultures. Prenatally administered antibiotics did not affect time to positivity in positive cultures drawn on the first day of life. In a selected group of microorganisms that are the frequent cause of bacteremia in term infants, 97% and 99% of cultures were positive by 24 to 36 hours of incubation when only pretherapy cultures are evaluated., Conclusions: The ESP blood culture system identified 77%, 89% and 94% of all microorganisms at 24, 36, and 48 hours of incubation in aerobic cultures obtained from both term and preterm infants. Introduction of antimicrobial therapy did not affect time to positivity. Reducing duration of antibiotic therapy to 24 to 36 hours should be considered in term, asymptomatic newborn infants undergoing evaluation for suspected sepsis for maternal indications. Confirmation of similar rapidity of detection using other blood culture systems should be undertaken.

A prospective study in pediatric patients compared the BACTEC system, an automated blood culture system using Bactec Ped Plus/F broth media, with the conventional system using Brain Heart Infusion broth media. Each single blood sample drawn for culture from hospitalized children was evaluated by both systems simultaneously. Of 244 pairs of blood samples, 44 (18%) were positive for microorganisms. Of these isolates, 24 (55%) were detected by both systems, 10 (23%) were detected by the BACTEC system only, and 10 (23%) were detected by the conventional system only. The mean turn around time of the BACTEC system (0.56 +/- 0.34, range 0.08-1.55 days) was significantly shorter than that of the conventional system (3.36 +/- 2.72, range 1-7 days, p < 0.001). Seven isolates strongly suspected to be due to contaminants grew out after 5 days of incubation and were detected by the conventional system only. In this study the BACTEC system and the conventional system were equally effective in detecting microorganisms in the patients' blood samples even if antibiotic therapy had been previously administered. However, the results from the BACTEC system were available much sooner and less likely to be contaminants.

The newly developed Hemofast MRSA system for Staphylococcus aureus identification and methicillin-resistant staphylococci (MRS) detection in blood culture broths was evaluated in 106 Bactec broths containing grapelike clusters of Gram-positive cocci. All 26 S. aureus positive broths were correctly identified by Hemofast MRSA within two hours, and 81% were identified within one hour. Sensitivity and specificity were both 100%. Accuracy of MRS detection was 100%, i.e., no discrepancies versus the agar diffusion method were found. When the 28 broths containing coagulase negative staphylococci (CNS) were tested using an inoculum ten fold larger than for S. aureus testing, a number of discrepancy were recorded. For 49 other broths containing CNS, accuracy was 98.5% when the test was interpreted based on the growth rate of the strain, according to the manufacturer's instructions. One broth containing S. epidermidis strain susceptible yielded a false result with Hemofast MRSA, indicating that it probably contained a contaminant. The other discrepancies occurred with specimens containing mixed populations with CNS or a Micrococcus strain. Most (85%) results were available within 4 hrs 30 min, irrespective of the S. aureus or CNS strains. Avoiding the isolation step on agar, Hemofast MRSA saves 24 to 48 hours, thus allowing earlier antistaphylococcal treatment.

Background: Concentration of microorganisms in blood is low in bacteremia from extravascular sources. The best yield from blood cultures is achieved by culturing a minimum of 10-20 ml, although in some processing blood culture systems such as Bactec NR-860, smaller volume is used. The objectives of the present study were to establish the frequency in which inadequate small blood volumes are employed for culturing and to analyze the relation between the cultured blood volume in Bactec 6A bottles and the yield achieved., Material and Methods: We weighed 2000 Bactec 6A bottles pertaining to consecutive blood cultures obtained from untreated patients with clinical suspicion of Infection. The cultured blood volume was estimated subtracting the mean empty bottle weight., Results: Microorganisms were recovered from 251 bottles (12.5%). One hundred and thirty one (6.8%) isolates were considered as clinically significant and 115 (5.7%) as contaminant. The inoculated blood volume in both significant (5.532 +/- 1.587 ml) and non-significant (5.471 +/- 1.563 ml) recoveries was superior than that of bottles without microbiologic growth (5.209 +/- 1.575 ml, p = 0.016 and p = 0.06, respectively). A linear positive trend was found between the cultivated blood volume and the rate of recoveries (p = 0.008). Within the range of 1 up to 10 ml, the rate of recoveries increased 2.28% for each additional ml of cultivated blood (r = 0.953, p < 0.0001). Out of the 2,000 weighed bottles 127 (6.3%) contained less than 3 ml of blood and 576 (29%) between 3 and 5 ml., Conclusions: We have proved that the rate of recoveries from Bactec 6A bottles increased with the volume of cultured blood. In untreated patients, this increase is maintained up to volumes of 7 to 10 ml.

Standard references continue to recommend testing specimens processed on the BACTEC TB System for 8 weeks, despite evidence that mycobacteria are rarely recovered beyond 5-6 weeks. To clarify this issue, we retrospectively reviewed all positive blood/bone marrow cultures processed during a 17-month period when specimens were tested for 6 weeks. We then prolonged the incubation period to 8 weeks during the subsequent 5 months. Excluding Mycobacterium genavense, only 1 of 159 mycobacterial isolates was recovered during and none were recovered beyond the 5th week of incubation and testing on the BACTEC TB System.

The BacT/Alert (BTA) (Organon Teknika, Durham, NC) and Isolator 10 (ISO) (Wampole Laboratories, Cranbury, NJ) blood culture systems were evaluated for their ability to detect aerobic and facultatively anaerobic microorganisms in blood of adult patients. For each culture 8 mL of blood was inoculated into both the aerobic standard BTA bottle and the ISO tube. Of 7,259 paired culture sets, 1,168 organisms were recovered, and 667 (57.1%) of these were considered clinically significant. This represented 540 clinically significant positive cultures from 266 patients. Of the significant isolates, 410 were recovered by both systems, 108 by BTA only and 149 by ISO only (P <.025). Overall, the BTA detected 77.7% of the significant isolates, whereas ISO detected 83.8%. The ISO recovered significantly more isolates of Staphylococcus aureus (P = .0001), coagulase-negative Staphylococcus spp (P <.01), and non-Enterobacteriaceae gram-negative rod species (P <.0025), whereas the BTA detected significantly more isolates of Streptococcus spp (P <.0025). Growth of S aureus (P <.0025), Enterococcus spp (P <.0025), and Streptococcus spp (P <.0075) was detected earlier by the BTA when laboratory coverage was available during the first shift only (7:30 AM to 4:00 PM), and additionally of Enterobacteriaceae (P <.0005) and other gram-negative rod species (P <.0001) if coverage was extended to 12:00 AM. Yeasts were detected more rapidly by the ISO (P <.0025). The ISO contamination rate (5.9%) was six times that of the BTA. Taking into account its ability to rapidly detect most organisms, its automated and thus labor-saving features, and the minimal contamination rate associated with its use, the BTA appears to be a reliable alternative to the ISO as a blood culturing system, although improvement in detection of staphylococci and non-Enterobacteriaceae gram-negative rods would be desirable.

The ESP 80A aerobic blood culture of the ESP automated blood culture system (Difco Laboratories. Detroit, Mich.) was compared with two manual aerobic blood culture systems, the Isolator (Wampole Laboratories, Cranbury, N.J.) and the Septi-Chek (Becton Dickinson, Cockeysville, Md.) systems, for the detection of bloodstream microorganisms from 5,845 blood samples for culture collected from adult patients with suspected septicemia. The bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated onto solid medium and this medium was incubated for 72 h. A total of 609 microorganisms were recovered from 546 blood cultures. There was no statistically significant difference in the total recovery of microorganisms for the ESP 80A system when compared with that for the Septi-Chek system (P = 0.083); however, the Isolator system recovered significantly more microorganisms overall than either the ESP 80A (P < 0.001) or the Septi-Chek (P < 0.001) system. When assessing individual probable pathogens, the Isolator system detected statistically significantly more Staphylococcus aureus and Candida spp. than either the ESP 80A or the Septi-Chek system (P < 0.05). Similarly, the Isolator system detected statistically significantly more bloodstream infections (septic episodes) caused by S. aureus and Candida spp. than either the ESP 80A or the Septi-Chek system (P < 0.05). In blood culture sets which produced growth of the same probable pathogens in the ESP 80A and the Isolator systems, there was no statistically significant difference in the median times to detection for all pathogens combined (P = 0.067). However, a similar comparison showed the Isolator and the ESP 80A systems to have statistically significantly shorter median detection times for all pathogens combined (P < 0.001) when they were independently compared with the Septi-Chek system. The ESP 80A system had 29 (0.5%) false-positive signals. The ESP system required less processing time than the Isolator system and eliminates the hands-on time for the detection of positive cultures required by the manual systems.

One reason for the underdiagnosis of bacteremia as an etiology of fever in neutropenic patients might lie in inefficient blood culture techniques. This possibility was investigated in a retrospective study by comparing the efficacies of a manual and an automated blood culture system used to detect bacteremia in such patients. All neutropenic fever episodes accompanying all 93 intensive chemotherapy cycles of 26 consecutive patients with acute myelogenous leukemia (AML) enrolled in Finnish Leukemia Group AML 86 trial and treated in Oulu University Hospital over 3 1/2 years were analyzed. The chemotherapy protocol and the supportive care of the patients remained the same during the whole period. In 1990-91 the blood cultures were made manually and in 1992-93 with an automated continuous-monitoring culture screening system. Evaluable febrile episodes numbered 53 during 1990-91 and 73 during 1992-93. There was a statistically significant increase (p < 0.05) of culture-positive episodes, from 21% to 40%, when the continuous-monitoring system was adopted. The new method proved to be better than the manual one in detecting bacteremia of neutropenic patients.

The BACTEC 9240 (Becton Dickinson, Sparks, Md.) automated blood culture system is based on the continuous monitoring of CO2 production by means of a fluorescent sensor attached to the bottom of culture vials. We compared the performance of the BACTEC aerobic Plus/F medium to that of the Septi-Chek Release medium (Becton Dickinson, Sparks, Md.), a manual biphasic blood culture system. Sets consisting of BACTEC aerobic Plus/F and Septi-Chek Release vials inoculated with similar volumes (7 +/- 2 ml) were evaluated. In the laboratory, systems were equipped and analyzed according to the manufacturer's recommendations. The BACTEC and Septi-Chek vials were incubated at 35 degrees C for 5 days. A total of 6,116 compliant sets were obtained from 1,972 adult patients (3.1 cultures per patient). Of these, 731 (12%) were culture positive, including 612 (10%) that yielded at least one pathogen, and 143 (2%) were considered to be contaminated. Of the 672 pathogenic organisms detected, 524 were isolated from the BACTEC aerobic Plus/F medium and 574 were isolated from the Septi-Chek Release medium, 428 organisms grew in both media, 96 organisms grew only in the BACTEC aerobic Plus/F medium, and 146 organisms grew only in the Septi-Chek Release medium (P = 0.001). Haemophilus spp. were isolated more often (P = 0.03) from the BACTEC aerobic Plus/F medium; however, more Streptococcus anginosus organisms (P = 0.02), members of the Enterobacteriaceae family (P < 0.03), and gram-negative anaerobes (P = 0.03) were isolated from the Septi-Chek Release medium. Pathogenic organisms were detected significantly earlier (P < 0.0001) with the BACTEC aerobic Plus/F medium in conjunction with the BACTEC 9240 instrument than with the Septi-Chek Release medium.

A controlled clinical comparison was carried out with the BACTEC 9240 Aerobic/F resin bottle and the Isolator system with adult patients suspected of having bloodstream infections. A total of 10,500 paired specimens were collected, of which 1,122 from 520 patients were positive. There were 68 false-positive BACTEC bottles; 259 positive cultures that were excluded from analysis because the bottle, the Isolator, or both failed to meet the minimum volume criterion of 8 ml of blood; and 207 positive cultures that were excluded because the isolates were found to be clinically insignificant or of indeterminate clinical significance on the basis of patient assessment. A total of 656 positive cultures from 258 patients formed the basis of the analysis. Significantly more Staphylococcus aureus isolates (P = 0.03), Staphylococcus epidermidis isolates (P = 0.03), members of the family Enterobacteriaceae (P = 0.03), and Pseudomonas aeruginosa isolates (P = 0.04) were recovered from the resin bottle, and there was no category of organism that was recovered significantly more frequently from the Isolator system. With patients receiving antibiotics at the time of blood culture, S. aureus, S. epidermidis, and gram-negative bacilli were recovered significantly more frequently from the resin bottle. No significant differences between systems were found with cultures from patients not receiving antibiotics at the time of blood culture. Only 12 clinically significant organisms were recovered from the bottle on terminal subcultures, and only 1 of these had not been previously isolated from another blood culture set (10 of the 12) or from the companion Isolator (1 of 12). The Aerobic/F resin bottle continuously monitored in the BACTEC 9240 instrument proved to be superior to the Isolator in overall yield of organisms causing bloodstream infection in adults and required less technician time for specimen processing and examination than the Isolator system.

The performance characteristics of the Isolator (Wampole Laboratories, Cranbury, N.J.) and the BacT/Alert (Organon Teknika Corporation, Durham, N.C.) aerobic blood culture systems were compared for 6,009 blood culture sets obtained from patients with suspected bloodstream infections. The BacT/Alert aerobic bottle [BTA(O2)] was continuously agitated while it was incubated in 5% CO2 at 36 degrees C; culture plates prepared from the Isolator tube [I(O2)] were incubated in 5% CO2 at 37 degrees C. From 394 blood cultures, 416 clinically significant isolates of bacteria and yeasts were recovered. The overall yields for BTA(O2) and I(O2) were not significantly different (319 versus 336; P = 0.20). I(O2) recovered significantly more staphylococcus (P < 0.05) and yeast isolates (P < 0.01). BTA(O2) recovered significantly more aerobic and facultatively anaerobic gram-negative bacilli (P < 0.05). In blood culture sets which produced growth of the same organisms in both the BTA(O2) and I(O2) systems, the BTA(O2) system detected growth sooner, but more rapid identification was possible with the I(O2) system by virtue of earlier isolation of colonies on solid media.

In a study comparing the BACTEC 9240 (Becton Dickinson, Canada Inc., Mississauga, Ontario, Canada) and the BacT/Alert (Organon Teknika, Scarborough, Ontario, Canada) blood culture systems, 6,437 sets of four bottles of 10,412 cultures submitted were deemed acceptable for evaluation. There were 586 clinically significant isolates from 544 positive blood cultures. Of the 544 positive blood cultures, 329 were positive in both systems, 157 were positive in the BACTEC 9240 system alone, and 58 were positive in the BacT/Alert system alone (P < 0.001). These cultures represented 421 bacteremic episodes, of which 290 were detected by both systems, 84 were detected by the BACTEC 9240 system alone, and 47 were detected by the BacT/Alert system alone (P < 0.001). The difference between the two systems in terms of microbial recovery was attributable largely to differences in the ability to grow staphylococci. Of 82 monomicrobial isolates of Staphylococcus aureus, 46 were isolated by both systems, 30 were isolated by the BACTEC 9240 system alone, and 6 were isolated by the BacT/Alert system alone (P < 0.0001). A similar pattern was noted in the case of coagulase-negative staphylococci, of which 48 were isolated in both systems, 46 were isolated in the BACTEC 9240 system alone, and 5 were isolated in the BacT/Alert system alone (P < 0.0001). The false-positive rate in the BACTEC 9240 system was 0.5%, and that in the BacT/Alert system was 1.2%. Both systems provided satisfactory service mechanically, and in terms of computer software and costs, both systems were similar. The differences in the recovery of organisms, notably staphylococci, may be due in part to the fact that in the present study the BacT/Alert medium did not contain antimicrobial agent-removing devices, whereas the BACTEC 9240 medium did. Until this difference is resolved, we consider the BACTEC 9240 system to more adequately meet our hospital's requirements.

The Organon Teknika BacT/Alert (Organon Teknika, Durham, NC), using the Pedi-BacT 20 mL aerobic bottle (BPBCS) was compared to the Wampole Isolator (WI) 1.5 Microbial tube (Wampole Laboratories, Cranbury, NJ), for detection and recovery of pediatric pathogens. The BPBCS continuously monitors culture bottles for changes in CO2 concentrations, while WI cultures are examined twice daily for appearance of colonial growth on agar media. Of 5,175 paired blood cultures, 383 pathogens were recovered from 606 positive cultures. There were 272 pathogens recovered by both systems, 64 from BPBCS only, and 47 from WI only. Overall recovery rates were 88% for BPBCS and 83% for WI. There was no significant difference between the two systems in detection or times to positivity of staphylococci, Enterobacteriaceae, or pseudomonads. Trends toward better recovery of streptococci (20 vs. 10) and fastidious microaerophiles (3 vs. 0) were found with BPBCS, whereas more slowly growing pathogens (Rochalimaea henselae [1], Mycobacterium avium-intracellulare [1]) were recovered by WI only, but because of their lower frequency did not achieve statistical significance. Detection of Haemophilus influenzae (14.9 hours in WI vs. 45.4 hours in BPBCS) was faster with WI. False positive plus contaminant cultures were detected in 5.9% BPBCS versus 1.5% WI. BPBCS offers detection of bacteremia at a rate comparable to WI with advantages of automation.

Bottles developed for use in the BacT/Alert automated blood culture system (Organon Teknika Corp., Durham, N.C.) can accept up to 10 ml of blood without falling below a 1:5 ratio of blood to broth. We compared the yield and speed of detection of microorganisms in 13,128 adequately filled, paired, aerobic bottles inoculated with 5 versus 10 ml of blood at three university hospitals. A total of 798 microorganisms causing sepsis grew in one or both bottles. The overall recovery of microorganisms from 10-ml samples exceeded that from 5-ml samples (P < 0.001); the increased yield attributed to the additional 5 ml in the samples was 7.2%. The increased yield from 10-ml inocula was most marked for Escherichia coli (P < 0.01) and other members of the family Enterobacteriaceae (P < 0.001). Ten-milliliter samples did not yield more gram-positive bacteria, nonfermentative gram-negative rods, or yeasts. When both bottles were positive, the bottles inoculated with 10 ml of blood showed growth sooner (P < 0.001). Earlier detection with 10-ml inocula was especially notable for coagulase-negative staphylococci (P < 0.001), streptococci (P < 0.001), E. coli (P < 0.025), and other members of the family Enterobacteriaceae (P < 0.025). We conclude that an increase in the volume of blood inoculated into BacT/Alert aerobic blood culture bottles from 5 to 10 ml will increase the overall yield and the speed of detection of clinically important blood pathogens.

The recovery of pathogens and the speed of their detection were determined for our conventional blood culture system (an Isolator [Wampole] and a 100-ml Thiol bottle [Difco]) compared with automated ESP aerobic and anaerobic bottles (80 ml each; Difco). Each of the four culture devices was inoculated with approximately 10 ml of blood from symptomatic patients weighing more than 80 lb (ca. 36 kg). From 7,070 sets of cultures for 2,841 patients, 607 clinically significant isolates were recovered: 456 (75.1%) from the Isolator, 353 (58.2%) from Thiol, 377 (62.1%) from ESP aerobic bottles, and 346 (57.0%) from ESP anaerobic bottles. Of the 607 isolates, 149 (24.5%) were detected only with the conventional system (Isolator and/or Thiol), and 65 (10.7%) were detected only with the ESP two-bottle system (P < 0.001). Our conventional system allowed for detection of significantly more isolates of members of the family Enterobacteriaceae (P < 0.001), Staphylococcus aureus (P < 0.01), Staphylococcus spp. (coagulase-negative) (P < 0.01), and Enterococcus spp. (P < 0.05), and ESP facilitated detection of significantly more isolates of S. pneumoniae (P < 0.01). When all four devices in a culture set were positive for the same isolate, no microbial species or group was detected significantly earlier ( > or = 24 h) by either blood culture system. The Isolator contamination rate (4.8%) was > or = 6 times the rate for any of the bottles. Of pathogens detected by the Isolator, 50% were recovered in counts of < or = 1.0 CFU/ml and 18% were recovered only as a single colony. The ESP system offered an automated, less labor-intensive blood culture system for which routine subcultures were not required, but the important considerations of culturing large volumes of blood and of obtaining at least two sets from each patient in our population were reemphasized.

The Difco ESP and Organon Teknika BacT/Alert (BTA) systems were evaluated in a clinical study of 5,421 aerobic and 5,035 anaerobic blood cultures. Of 405 clinically significant positive cultures evaluated, 272 grew in both systems, 86 grew in ESP only, and 47 grew in BTA only (P < 0.005). Of 320 organisms detected in aerobic bottles, 208 grew in both systems, 68 grew in ESP only and 45 grew in BTA only (P < 0.05), with Staphylococcus aureus the only organism showing a statistically significant difference. The ESP anaerobic bottle also detected more anaerobes (16 of 17 versus 4 of 17, P < 0.005) and more organisms overall (57 versus 34, P < 0.05). However, with the exception of patients with anaerobic bacteremia (12 of 13 for ESP and 4 of 13 for BTA, P < 0.05), there was no statistical difference in the detection of patient episodes. Average detection time of matched aerobic bottles was 18.3 h for ESP and 22.0 h for BTA (P < 0.001). For matched pairs of anaerobic bottles, the average detection time was faster in the BTA bottles (P < 0.001), because of the growth of facultative organisms. To explore the differences in anaerobic detection more fully, 20 sets of anaerobic bottles were seeded with 12 anaerobic species mixed with human blood. ESP grew more organisms (17 of 20 versus 10 of 20, P < 0.025), and the average time to detection for the 10 paired positive cultures was 21.6 h for ESP and 50.8 h for BTA (P < 0.05). Times for loading and unloading bottles were similar for both systems.

In the midst of technologic advances within blood culture microbiology, several manual blood culture systems, which have an important role in the detection of bacteremia, mycobacteremia, and fungemia, are often overlooked. These include traditional broth-based systems, agar-broth biphasic blood culture techniques, a commercial broth-based manual system in which growth is detected by a broth displacement method, and lysis-centrifugation. This article reviews the operational features, advantages, and disadvantages of each.

For the first time, clinical microbiologists have the option of purchasing or leasing a variety of automated blood culture systems. Although the performance characteristics of the BACTEC radiometric and nonradiometric blood culture systems are well-documented, we are only beginning to learn the performance characteristics of CMBCS. Well-designed and executed controlled clinical trials are needed to establish performance characteristics and to address issues that are unique to CMBCS. In particular, controlled clinical comparisons of different CMBCS are needed. In terms of the impact on clinical microbiology laboratories, there are no published data to support claims that the use of these systems decreases laboratory workload, staffing, or costs. Although such an impact seems reasonable and even likely, unequivocal documentation of such savings would make it easier for many laboratorians to justify the cost of purchasing or leasing one of these systems. Similarly, there are no published data regarding the long-term reliability of these systems while in clinical use. A documented track record of mechanical and electrical reliability would be useful to clinical microbiologists contemplating the acquisition of a new blood culture system.

Rapid methods of determining microbial contamination are needed in suspecting contaminated banked blood or other blood products. In this study, we experimented contaminated blood units with 122 strains of bacteria and fungi. After innoculation, a comparison between ESP Blood Culture System (Difco Laboratories Inc., Detroit MI), BacT/Alert (Organon Teknica, Durham, NC) and Bactec 9240 System was made for their efficiency in the detection of microbial contamination. Experimental data showed a diagnostic relevance of these automated systems with no significant differences in time detection of microbial contamination between the three methods under comparison.

A multipoint inoculation method was used for the terminal subculture of blood cultures. This economical yet reliable technique successfully isolated important human pathogens such as Haemophilus influenzae from macroscopically negative blood cultures.