Paired Blood Cultures Research Articles

Utility of paired blood cultures and smears in diagnosis of disseminated Mycobacterium avium complex infections in AIDS patients.

For 273 patients evaluated for disseminated Mycobacterium avium complex infection, a total of 1,047 mycobacterial blood cultures (MBCs) were submitted; the M. avium complex was recovered from 140 (13%) of the specimens. Results for the paired MBCs were highly concordant: in 392 of 462 (85%) culture sets, both MBCs were negative, in 53 of 462 (11%) sets, both MBCs were positive, and in only 17 of 462 (4%) sets was one culture positive and the other negative. Acid-fast smears were done on sediments from 671 specimens; smears were positive for 4 of 98 (4%) cultures that grew the M. avium complex. A single MBC should be obtained and then repeated if negative and disseminated M. avium complex infection is still clinically suspected. Use of direct acid-fast smears of sediments is not a reliable means of detecting mycobacteremia.

Effect of agitation of BACTEC 13A blood cultures on recovery of Mycobacterium avium complex

The effect of agitation of BACTEC 13A bottles (Becton Dickinson) on the recovery of Mycobacterium avium complex (MAC) from blood was compared with that of static incubation. A total of 265 blood specimens was inoculated in duplicate into BACTEC 13A bottles. One specimen was statically incubated at 35 degrees C, and the other was incubated with agitation on a Gyrotory shaker at 35 degrees C for the first 2 weeks and thereafter without shaking for up to 12 weeks. Of the 265 specimens, 77 (29.1%) were positive in either one or both of the paired bottles. The average detection times for the shaken and nonshaken bottles were 12.7 and 15.9 days, respectively. A total of 10.4% of the specimens in the shaken bottles became positive 1 week before those in the nonshaken bottles, and 16.9% of the shaken cultures were positive more than 2 weeks before their counterparts. A further 46.8% of the agitated specimens became positive while the corresponding nonagitated cultures remained negative. When both specimens became positive at the same time, 88% of the shaken cultures had higher growth indices than their nonshaken counterparts. A further 11 paired blood cultures were taken from patients known to be infected with MAC to assess the effect of agitation of bottles on the utility of making twice-weekly readings during the first 2 weeks of incubation. Ten of the 11 sets of specimens in the shaken bottles were positive 1 or more weeks before those in the corresponding nonshaken bottles. In the remaining set, both specimens became positive on the same day; however, the growth index of the agitated culture was higher.(ABSTRACT TRUNCATED AT 250 WORDS)

Controlled evaluation of BACTEC Plus 26 and Roche Septi-Chek aerobic blood culture bottles

Blood culture bottles that accept up to 10 ml of blood have recently been introduced for the BACTEC nonradiometric blood culture system. The new formulation, designated BACTEC Plus, contains 25 ml of tryptic soy broth, 0.05% sodium polyanetholesulfonate, and mixed resins. In a collaborative study conducted at three university hospitals, we evaluated the BACTEC Plus 26 (BP26) aerobic bottle and the Roche Septi-Chek aerobic bottle with its agar slide paddle in 5,293 paired blood cultures. Significantly more microorganisms (P less than 0.001), especially Staphylococcus aureus (P less than 0.001), Staphylococcus epidermidis (P less than 0.01), enterococci (P less than 0.005), and members of the family Enterobacteriaceae (P less than 0.005), were detected by the BP26 bottle. When both bottles detected growth, BP26 did so earlier (P less than 0.001). In particular, S. epidermidis (P less than 0.001), streptococci (P less than 0.005), enterococci (P less than 0.05), and members of the family Enterobacteriaceae (P less than 0.001) were detected earlier by the BP26 bottle. We conclude that the BP26 bottle provides a yield and speed of detection of microorganisms superior to those of the Roche Septi-Chek aerobic blood culture bottle.

Clinical comparison of isolator and BACTEC 660 resin media for blood culture

The 10-ml Isolator system (E.I. du Pont de Nemours & Co., Inc., Wilmington, Del.) was compared with the BACTEC 16A-17A nonradiometric resin system (Johnston Laboratories, Inc., Towson, Md.) for isolation of organisms from 6,839 paired blood cultures. Equal volumes of blood (6 to 10 ml for each Isolator and 3 to 5 ml for each BACTEC bottle) were cultured in parallel in the two systems, and 600 isolates that were judged to be clinically significant by chart review were recovered during the study. The BACTEC resin system detected 510 (85%) and the Isolator system detected 435 (72%) of the clinically significant isolates (P less than 0.001). Of 45 polymicrobial blood cultures, the BACTEC system detected 32 (71%) and the Isolator system detected 21 (47%) (P less than 0.05). Of 253 gram-negative bacilli isolated during the study, 30% were detected only in the BACTEC system and 16% were detected only in the Isolator system (P less than 0.001), and of 56 nonfermentative or fastidious gram-negative bacilli detected, 46% were recovered only in the BACTEC system, while 14% were detected only in the Isolator system (P less than 0.001). Of 86 streptococci isolated during the study, 30% were detected only in the BACTEC system, and 4% were detected only in the Isolator system (P less than 0.001). Recoveries of anaerobic bacteria, staphylococci, and yeasts were equivalent in the two systems. Organisms judged to be contaminants were detected in approximately 1% of the cultures in each system. The results suggest that use of resin media renders the BACTEC nonradiometric system equivalent or superior to the Isolator system for detection of clinically significant organisms in blood cultures.

The use of bone marrow culture for the diagnosis of melioidosis

We have evaluated prospectively the contribution of bone marrow culture to the diagnosis of melioidosis. Bone marrow (BMC) and blood cultures (BC) were collected concurrently from 105 patients with suspected acute, severe melioidosis. 67 patients were subsequently proved to have the disease whilst other significant organisms were isolated from these specimens in 5 cases. Overall, 67·2% of BC and 64·2% of BMC from melioidosis patients grew Pseudomonas pseudomallei. Time to positivity did not differ significantly in paired BC and BMC specimens. These results do not support the routine use of BMC in the diagnosis of acute, severe melioidosis. In one patient with pulmonary melioidosis, however, blood cultures were repeatedly negative, whilst bone marrow grew P. pseudomallei, and this preceded the development of a distant focus of infection. This suggests that culture of bone-marrow may be of value in certain blood culture-negative patients with melioidosis.

Comparative evaluation of nonradiometric BACTEC and improved oxoid signal blood culture systems in a clinical laboratory

The BACTEC NR660 blood culture system, which uses infrared spectroscopy to detect carbon dioxide generated by bacterial growth, was compared with the new medium formulation of the Oxoid Signal system. Two trials were conducted: a comparative study of 88 organisms in simulated blood cultures and a clinical trial of 3,321 paired patient blood culture samples. Both trials showed that overall the BACTEC system performed better in the recovery of organisms. The Oxoid system was unable to detect by signal the growth of the majority of yeasts, nonfermentative gram-negative bacilli, Neisseria meningitidis, Nocardia spp., and Corynebacterium jeikeium. There were no significant differences in the yield of Staphylococcus spp., members of the family Enterobacteriaceae, Streptococcus spp., or anaerobic organisms. BACTEC detected growth more quickly than did the Oxoid system; 61% of the isolates were detected by BACTEC at 24 h, while 49% of the isolates were detected by Oxoid. The Oxoid system had a high proportion (58.5%) of false-positives, compared with 7.7% for the BACTEC system. Despite the new medium formulation of the Oxoid system, its performance is still not equivalent to that of the BACTEC system.

Quantitative aspects of septicemia

For years, quantitative blood cultures found only limited use as aids in the diagnosis and management of septic patients because the available methods were cumbersome, labor intensive, and practical only for relatively small volumes of blood. The development and subsequent commercial availability of lysis-centrifugation direct plating methods for blood cultures have addressed many of the shortcomings of the older methods. The lysis-centrifugation method has demonstrated good performance relative to broth-based blood culture methods. As a result, quantitative blood cultures have found widespread use in clinical microbiology laboratories. Most episodes of clinical significant bacteremia in adults are characterized by low numbers of bacteria per milliliter of blood. In children, the magnitude of bacteremia is generally much higher, with the highest numbers of bacteria found in the blood of septic neonates. The magnitude of bacteremia correlates with the severity of disease in children and with mortality rates in adults, but other factors play more important roles in determining the patient's outcome. Serial quantitative blood cultures have been used to monitor the in vivo efficacy of antibiotic therapy in patients with slowly resolving sepsis, such as disseminated Mycobacterium avium-M. intracellulare complex infections. Quantitative blood culture methods were used in early studies of bacterial endocarditis, and the results significantly contributed to our understanding of the pathophysiology of this disease. Comparison of paired quantitative blood cultures obtained from a peripheral vein and the central venous catheter has been used to help identify patients with catheter-related sepsis and is the only method that does not require removal of the catheter to establish the diagnosis. Quantitation of bacteria in the blood can also help distinguish contaminated from truly positive blood cultures; however, no quantitative criteria can invariably differentiate contamination from bacteremia.

Comparison between Bactec Nr. 660® and a conventional 12‐tube blood culture system

The detection power of the automated blood culture system Bactec NR 660, based on infrared detection of carbon dioxide in an agitated aerobic medium and a non-agitated anaerobic medium, was compared with that of our conventional 12-tube blood culture system. Of 1685 paired blood cultures, 258 (15.3%) were positive in one or both systems. Clinically relevant isolates were found in 11.5%. The dominating species were Escherichia coli(41%), followed by Staphylococcus aureus(14%) and Klebsiella spp.(8%). The Bactec system detected 178 (10.6%) and the 12-tube system 157 (9.3%) clinically relevant microorganisms after seven days' incubation. Significantly more clinically relevant isolates were detected by the Bactec system alone as compared with the conventional system alone (40 versus 19, p less than 0.01). The detection time was significantly shorter in the Bactec system for all isolates and for E. coli and S. aureus separately (p less than 0.01). 1.8% of the isolates in the Bactec system and 2.1% in the 12-tube system were considered clinically non-relevant contaminants.

Investigation of a Streptococcus viridans Pseudobacteremia Epidemic at a University Teaching Hospital

AbstractFor several months in 1986, an outbreak of Streptococcus viridans pseudobacteremia occurred at a large teaching hospital. All sources of laboratory blood culture contamination were excluded. A retrospective epidemiological study indicated that one phlebotomist, “P,” collected a disproportionate number of the positive blood cultures. Further comparison of the paired blood culture results from the three months when the incidence was highest revealed a good concordance of results among all other phlebotomists (Kappa = 0.5), while P's results concurred with others less frequently than would be expected even by chance (Kappa < 0.0). Clinical follow-up showed that P did not routinely wear gloves while drawing blood and had eczema of the hands. Skin scrapings from the hands, right index finger/fingernail grew predominantly S viridans species that were compatible with those recovered from contaminated blood cultures. This epidemic demonstrated the need for early detection of this source as a cause of nosocomial pseudobacteremia.

Investigation of a Streptococcus viridans pseudobacteremia epidemic at a university teaching hospital.

For several months in 1986, an outbreak of Streptococcus viridans pseudobacteremia occurred at a large teaching hospital. All sources of laboratory blood culture contamination were excluded. A retrospective epidemiological study indicated that one phlebotomist, "P," collected a disproportionate number of the positive blood cultures. Further comparison of the paired blood culture results from the three months when the incidence was highest revealed a good concordance of results among all other phlebotomists (Kappa = 0.5), while P's results concurred with others less frequently than would be expected even by chance (Kappa less than 0.0). Clinical follow-up showed that P did not routinely wear gloves while drawing blood and had eczema of the hands. Skin scrapings from the hands, right index finger/fingernail grew predominantly S viridans species that were compatible with those recovered from contaminated blood cultures. This epidemic demonstrated the need for early detection of this source as a cause of nosocomial pseudobacteremia.

Comparative evaluation of the BCB Roche and Oxoid Signal blood culture systems

The agar-slide blood culture system BCB Roche was compared with the Oxoid Signal blood culture system using 2,266 paired blood cultures. A total of 271 (12%) paired sets were culture-positive, including 222 (9.8%) yielding pathogens associated with septicemia and 50 (2.2%) yielding likely contaminants. In the recovery of the total 235 isolates considered as pathogens, the BCB Roche system yielded 202 (85.9%), the Oxoid Signal 211 (89.8%); 178 (75.7%) were cultured by both systems. There was no statistically significant difference between the two systems in their sensitivity, contamination-rate and detection time, except for gram-positive organisms, which were detected earlier by the Oxoid Signal system. Both systems performed well and were easy to handle.

Comparison of Du Pont Isolator and Roche Septi-Chek for detection of fungemia

The rapid detection of fungemia in hospitalized patients is imperative, particularly for those who are immunocompromised. Our laboratory compared the Roche Septi-Chek with the Du Pont Isolator for the recovery of fungi from blood. Of 23,586 matched pairs of blood cultures, 199 were positive. The Isolator detected 178 (89.4%) and the Septi-Chek detected 119 (59.7%) of all positive isolates. The mean recovery time for the Isolator and Septi-Chek was 2.2 and 4.9 days, respectively. The Isolator detected fungemia earlier than the Septi-Chek did and was the only culture system positive in 83% of 53 patients, whereas the Septi-Chek system yielded the same results in only 13% of the patients. The Isolator provides a more rapid and sensitive method for the recovery of fungi from blood.

Controlled evaluation of Trypticase soy broth in agar slide and conventional blood culture systems.

A commercially available biphasic blood culture system that utilizes an attachable agar slide paddle and Trypticase soy broth (BBL Microbiology Systems, Cockeysville, Md.) was compared with a conventional Trypticase soy broth blood culture bottle in 6,867 paired blood cultures from adult patients. Both systems were inoculated with equal volumes of blood (5 ml) and incubated aerobically (vented) for 2 weeks. More clinically important bacteria and fungi, including Staphylococcus epidermidis, streptococci, Escherichia coli, Klebsiella spp., other Enterobacteriaceae, and Pseudomonas aeruginosa, were recovered from the biphasic system (P less than 0.001). In contrast, more anaerobic bacteria of importance were recovered in the conventional bottle (P less than 0.01). Staphylococci (P less than 0.001), gram-negative facultative and aerobic bacteria (P less than 0.001), and fungi (P less than 0.001) were detected 1 or more days earlier in the biphasic system, whereas pneumococci (P less than 0.05) were detected earlier in the conventional bottle. Of 603 clinically important microorganisms that grew in the biphasic system, 601 (99.7%) were detected by day 7 of incubation, but only 403 of 490 microorganisms (82.2%) were detected by day 7 in the conventional bottle. Overall, the biphasic system was superior to the conventional bottle. For optimal detection of anaerobic bacteremia, however, the biphasic system should be used in conjunction with a complementary anaerobic conventional bottle.

Evaluation of the BACTEC 16B Medium in a Cancer Center

The routine use of the new resin-containing BACTEC 16B (Johnston Laboratories, Inc., Cockeysville, MD) culture medium was evaluated in a population consisting primarily of cancer patients. Of 1,163 paired blood culture sets collected, 652 (56%) were collected in the presence of antimicrobial therapy. Eighty-three aerobic and facultatively anaerobic isolates were recovered from 79 positive blood culture sets. No significant difference could be demonstrated between the 16B and the 6B medium in the group of blood cultures collected from patients not receiving antibiotic therapy at the time of blood collection. In contrast, a significantly greater proportion of isolates (P less than 0.005) was recovered from the 16B medium (96%) than the 6B medium (68%) in the group of blood cultures collected in the presence of antimicrobial therapy. In this group, 43% of the isolates were either detected earlier or recovered solely from the 16B medium.

Coagulase-negative staphylococcal bacteremia in newborns.

Coagulase-negative staphylococci (C-NS) recently have been identified as common causes of septicemia in high-risk newborns. In this review of the incidence and clinical significance of C-NS blood culture isolates from infants with suspected septicemia in a newborn intensive care unit (NICU), 2.2 percent of blood cultures in an 18-month period yielded C-NS. Based on the criterion of growth in multiple blood cultures as evidence of bacteremia, six of 30 infants (20%) were bacteremic, 17 (57%) represented probable contaminants (on the basis of growth in only one of two paired blood cultures with CN-S), and seven cases (23%) were indeterminant (only one culture was obtained). In those infants considered bacteremic, onset occurred between 12 and 51 days of age. In those with probable contaminants, all but three occurred in the first week of life. All infants had manifestations of suspected sepsis and survived the clinical episode. These findings indicate that C-NS growth can be due to nosocomial bacteremia but frequently represents blood culture contamination, especially in infants in the first week of life. At least two paired peripheral blood cultures should be obtained in infants with suspected septicemia to differentiate bacteremia from contamination.

Evaluation of the BACTEC 16B medium in a cancer center.

The routine use of the new resin-containing BACTEC 16B (Johnston Laboratories, Inc., Cockeysville, MD) culture medium was evaluated in a population consisting primarily of cancer patients. Of 1,163 paired blood culture sets collected, 652 (56%) were collected in the presence of antimicrobial therapy. Eighty-three aerobic and facultatively anaerobic isolates were recovered from 79 positive blood culture sets. No significant difference could be demonstrated between the 16B and the 6B medium in the group of blood cultures collected from patients not receiving antibiotic therapy at the time of blood collection. In contrast, a significantly greater proportion of isolates (P less than 0.005) was recovered from the 16B medium (96%) than the 6B medium (68%) in the group of blood cultures collected in the presence of antimicrobial therapy. In this group, 43% of the isolates were either detected earlier or recovered solely from the 16B medium.

Controlled evaluation of hypertonic sucrose medium for detection of bacteremia and fungemia in supplemented peptone broth

Because the value of hypertonic media in detection of bacteremia and fungemia is controversial, we evaluated supplemented peptone broth (SPB) with 0.03% sodium polyanetholsulfonate with and without 10% sucrose in 5,439 paired blood cultures from adult patients. The aerobic atmosphere, 1:10 ratio of blood to broth, and methods for processing blood cultures were identical. Only cultures with adequate blood samples (greater than or equal to 4 ml) were compared statistically. More clinically important bacteria were recovered from SPB with sucrose (P less than or equal to 0.001), including Staphylococcus epidermidis, Enterobacteriaceae, and Bacteroidaceae. However, only one of nine isolates of Neisseria gonorrhoeae grew in SPB with sucrose. Staphylococci (P less than 0.001), Enterobacteriaceae (P less than 0.01), Pseudomonas aeruginosa (P less than 0.01), and yeasts (P less than 0.05) were detected 1 or more days earlier in SPB with sucrose. The effect of sucrose on blood cultures appears to be medium dependent, based on comparisons of our results with those of published reports.