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S Mirrett

Publications and source records attributed to S Mirrett.

At least 55 records · Page 3Linked to original sources

Controlled comparison of a new Becton Dickinson agar slant blood culture system with Roche Septi-Chek for the detection of bacteremia and fungemia.

Two commercially available agar slide blood culture systems, Septi-Chek (Roche Diagnostics, Div. Hoffmann-La Roche Inc., Nutley, N.J.) and VACUTAINER agar slant (Becton Dickinson [BD] Vacutainer Systems, Rutherford, N.J.), were compared in 5,628 paired blood cultures from adult patients. The systems were inoculated with equal volumes (5 ml) of blood. There was no statistically significant difference between the two systems in the overall recovery of clinically important microorganisms; however, significantly more Escherichia coli (P less than 0.05) organisms were detected in the Septi-Chek system. Concurrently, significantly more contaminants (P less than 0.001) also were present in Septi-Chek bottles. When microorganisms grew in both systems, there was no significant difference in the time to recovery of important organisms, except that anaerobes were detected more quickly in the BD system. This study suggests that the BD VACUTAINER agar slant is an acceptable alternative to the Septi-Chek system for routine blood cultures.

Bacteria↗

Effect of agitation and terminal subcultures on yield and speed of detection of the Oxoid Signal blood culture system versus the BACTEC radiometric system.

In an initial evaluation, we found the Oxoid Signal blood culture system inferior to the BACTEC radiometric system for detection of some microorganisms causing septicemia (M. P. Weinstein, S. Mirrett, and L. B. Reller, J. Clin. Microbiol. 26:962-964, 1988). To determine whether modified processing of the Oxoid Signal blood culture system could improve its yield and speed of detecting positive cultures relative to the BACTEC radiometric system, we agitated all Oxoid bottles during the first 24 to 48 h of incubation and performed aerobic and anaerobic subcultures of all Oxoid bottles negative after 7 days of incubation. These modifications improved the overall performance of the Oxoid system, particularly with regard to the yield of streptococci, members of the family Enterobacteriaceae, and Haemophilus, Neisseria, and Acinetobacter spp. The speed of detecting positive cultures also was improved, especially within the first 24 h of incubation. However, the BACTEC system still detected more positive cultures (P less than 0.005), especially of obligate aerobes such as Pseudomonas aeruginosa (P less than 0.05) and yeasts (P less than 0.005). The BACTEC system also detected positive cultures earlier than the Oxoid system (e.g., at 24 h of incubation, 70.5% of BACTEC positive cultures detected versus 62.1% of Oxoid positive cultures detected). Further modifications of the Oxoid system which might include a revised medium, additional processing modifications, altered headspace atmosphere, or a complementary second broth medium should be considered, since the system is attractive in concept and is easy to use in the clinical laboratory.

Aerobiosis↗

Evaluation of modified trypticase soy broth versus supplemented peptone broth in the detection of bacteremia and fungemia.

In vitro, 1.2% gelatin counteracts the inhibition of growth of bacterial species by sodium polyanetholsulfonate in blood culture media. Additionally, 1% yeast extract has been used to promote bacterial growth. We compared the performance of supplemented peptone broth and Trypticase soy broth, both of which contained sodium polyanetholsulfonate, gelatin and yeast extract. Trypticase soy broth with gelatin and yeast extract inhibited (p less than 0.001) and delayed growth, especially of gram-positive (p less than 0.01) and gram-negative (p less than 0.005) anaerobic bacteria. Although the recovery of organisms usually inhibited by sodium polyanetholsulfonate was similar in supplemented peptone and Trypticase soy broths, supplemented peptone broth clearly was superior in the recovery of other organisms commonly found in blood cultures.

Bacteria↗

Controlled evaluation of blood culture medium containing gelatin and V-factor-analog for detection of septicemia in children.

Both Neisseria meningitidis and Haemophilus influenzae are important isolates recovered in blood cultures from septicemic children. Sodium polyanetholsulfonate is present in most blood culture media and can inhibit the growth of certain bacteria, including N. meningitidis. The addition of gelatin to blood culture media neutralizes this inhibition. The growth of H. influenzae is enhanced by specific growth factors such as hemin and NAD. The addition of gelatin and V-factor-analog (a proprietary supplement for enhancing the growth of H. influenzae) might have a positive effect on the yield and on the speed of detection of septicemia in children. To evaluate this possibility, we did 4,565 paired comparisons of blood cultured in BACTEC 6B (aerobic) medium with and without the addition of both 1.2% gelatin and V-factor-analog. More aerobic and facultative bacteria grew in the 6B than in the 6B-gelatin-V-factor-analog medium (P less than 0.01). Only seven isolates of Neisseria spp. were recovered during this study period, with the 6B medium performing as well as the supplemented medium. When microorganisms grew in both bottles, they did so at the same time except for H. influenzae and Candida albicans. H. influenzae was recovered earlier from the 6B-gelatin-V-factor-analog bottle (P less than 0.01), with a mean time to detection of 8.5 h compared with 15.9 h for the 6B bottle. C. albicans was recovered earlier from the 6B bottle (P less than 0.02), with a mean time to detection of 34.9 h compared with 71.6 h for the 6B-gelatin-V-factor-analog bottle. We conclude that the 6B medium in its present formulation is superior to bB supplemented with gelatin and V-factor-analog.

Adolescent↗

Comparative evaluation of Oxoid Signal and BACTEC radiometric blood culture systems for the detection of bacteremia and fungemia.

The Oxoid Signal (Oxoid U.S.A., Inc., Columbia, Md.) blood culture system is a newly described, innovative method for visually detecting growth of microorganisms (D. Sawney, S. Hinder, D. Swaine, and E.Y. Bridson, J. Clin. Pathol. 39:1259-1263, 1986). We did 5,999 paired comparisons of equal volumes (10 ml) of blood in the Oxoid Signal and BACTEC (Johnson Laboratories, Towson, Md.) radiometric blood culture systems at two university hospitals that use identical methods of obtaining and processing specimens. Overall, more microorganisms were detected in the BACTEC system (P less than 0.001), in particular, streptococci (P less than 0.01), fungi (P less than 0.001), and nonfermentative gram-negative rods, especially Acinetobacter species (P less than 0.001). Trends favoring the BACTEC system for detection of Pseudomonas aeruginosa, Haemophilus species, and Neisseria species were noted. There were no differences in the yield of staphylococci, members of the family Enterobacteriaceae, and anaerobic bacteria. When both systems detected sepsis, the BACTEC did so earlier (P less than 0.001). This advantage was most notable at 24 h (70% of BACTEC positives detected versus 48% of Oxoid positives). The proportion of positives detected after 48 h, however, was similar (BACTEC, 84%; Oxoid, 78%). Revisions in the Oxoid Signal system itself or in the processing of Oxoid bottles appear to be necessary to improve its performance in detecting certain microorganism groups, especially fungi.

Bacteria↗

Comparative evaluation of medium and atmosphere of incubation for isolation of Streptococcus pyogenes.

To evaluate the effects of medium and atmosphere of incubation for the isolation of group A (GA) streptococci from throat cultures, we compared 1098 throat swabs plated on each of three blood agar plates (BAP). Two plates contained trimethoprim-sulfamethoxazole (SXT) and one had no antibiotics. The antibiotic-free medium and one SXT plate were incubated anaerobically (AnO2) at 35 degrees C, whereas the other SXT plate was incubated in air (O2) at 35 degrees C. The BAP-AnO2 identified 85% of 201 GA compared with 96% for SXT-AnO2 and 92% for SXT-O2. GA were confirmed from only 29% of the beta-hemolytic colonies on BAP-AnO2 compared with 53% from SXT-AnO2 and 44% from SXT-O2. SXT, however, appeared to delay recognition of GA; since 94% of positives were found on day 1 with BAP-AnO2 versus 80% and 62% with SXT-AnO2 and SXT-O2, respectively. We conclude that BAP-SXT (AnO2 or O2) is more sensitive and more specific than BAP-AnO2, but that a second day of incubation is required for optimal recovery of GA on the SXT selective media studied.

Aerobiosis↗

Controlled evaluation of trypticase soy broth with and without gelatin and yeast extract in the detection of bacteremia and fungemia.

The addition of gelatin to blood culture media has been suggested to prevent the inhibition of Neisseria meningitidis, Neisseria gonorrhoeae, Gardnerella vaginalis, and Peptostreptococcus anaerobius that is caused by sodium polyanetholsulfonate. To determine the effect of such supplementation on the overall yield of microorganisms, we compared the yield and speed of detection of clinically important microorganisms from 5422 paired 10-ml samples of blood cultured in Trypticase soy broth (TSB) containing 0.03% sodium polyanetholesulfonate (SPS) and TSB/SPS containing 1.2% gelatin and 1.0% yeast extract (mTSB). The atmosphere of incubation (open venting unit) and ratio of blood to broth (1:5) were the same for both samples. Only cultures with adequate blood sample (greater than or equal to 80% of stated volume) were compared statistically. Addition of gelatin and yeast extract resulted in inhibited growth of Enterobacteriaceae (p less than 0.001), Pseudomonas aeruginosa (p less than 0.01), fungi (p less than 0.05), and the overall set of microorganisms encountered (p less than 0.001). It delayed growth of Enterobacteriaceae (p less than 0.001) but reduced the time to recover staphylococci (p less than 0.02). Of 12 isolates of species usually inhibited by SPS, seven grew only with the addition of gelatin and yeast extract, none grew only without supplementation, and five grew in both media. Although gelatin and yeast extract may improve the yield of some specific bacteria, the routine use of these additives cannot be recommended for all blood culture media.

Adult↗

Controlled evaluation of modified radiometric blood culture medium supplemented with gelatin for detection of bacteremia and fungemia.

Although the addition of 1.2% gelatin to broth blood culture media containing sodium polyanetholesulfonate has been shown to enhance detection of certain bacteria, including Neisseria meningitidis, N. gonorrhoeae, Peptostreptococcus anaerobius, and Gardnerella vaginalis, the effect of such supplementation on the detection of other microorganisms causing bacteremia and fungemia is not known. Therefore, we studied BACTEC 6B medium with and without gelatin in 6,833 paired comparisons to examine the effects of supplementation on both the yield and the speed of detection of sepsis. More aerobic and facultative bacteria grew in the 6B than in the 6B-gelatin medium (P less than 0.001), especially staphylococci (P less than 0.01), Escherichia coli (P less than 0.01), other members of the family Enterobacteriaceae (P less than 0.05), and Acinetobacter spp. (P less than 0.05). When microorganisms grew in both bottles, they did so earlier in 6B than in 6B-gelatin (P less than 0.001). We conclude that the 6B medium in its present formulation is superior to 6B medium supplemented with 1.2% gelatin.

Bacteria↗

Controlled evaluation of the agar-slide and radiometric blood culture systems for the detection of bacteremia and fungemia.

A commercially available agar-slide blood culture bottle (Septi-Chek; Roche Diagnostics, Div. Hoffman-La Roche, Inc., Nutley, N.J.) was compared with the radiometric blood culture system (BACTEC; Johnston Laboratories, Inc., Towson, Md.) in 8,544 paired blood cultures from adult patients. The systems were inoculated with equal volumes (10 ml) of blood. Overall, there was no statistically significant difference between the two systems in the recovery of clinically important microorganisms, but significantly more members of the family Enterobacteriaceae other than Escherichia coli were detected by the agar-slide system (P less than 0.005). The agar-slide system detected more fungi, and the BACTEC detected more anaerobic bacteria; however, small numbers of recovered organisms precluded statistical significance. When microorganisms grew in both systems, their presence was detected one or more days earlier in the BACTEC (P less than 0.001). More contaminants grew in the agar-slide system (P less than 0.001). Both systems performed well, and either system should provide high yield and prompt detection of positive blood cultures in patients with bacteremia and fungemia if used in an optimal way as recommended by the respective manufacturers.

Bacteria↗

Controlled evaluation of supplemented peptone and Bactec blood culture broths for the detection of bacteremia and fungemia.

Comparison of conventional blood culture media with newer formulations of Bactec media for radiometric detection are lacking. Therefore, we compared the yield and speed of detection of clinically important microorganisms with supplemented peptone broth (SPB) and Bactec aerobic (6B) and anaerobic (7C or 7D) broths in 7,627 blood samples from adult patients. Acridine orange stains from SPB, radiometric readings from Bactec, and routine subcultures from all bottles were done at the same time intervals. Bactec grew more facultative gram-positive bacteria (P less than 0.02), Bacteroides spp. (P less than 0.001), gram-negative anaerobes (P less than 0.001). The two-bottle Bactec system required less time to detect Staphylococcus aureus (P less than 0.001), facultative gram-positive bacteria (P less than 0.001), Escherichia coli (P less than 0.02), facultative gram-negative bacteria (P less than .001), and fungi (P less than 0.001). Overall, Bactec yielded 11% more microorganisms and detected bacteremia sooner in 18% of samples than did SPB. This advantage was not because of radiometric monitoring, since most positive Bactec bottles were detected macroscopically. SPB offered no advantage for any group of microorganisms. We conclude that Bactec 6B and 7C or 7D broths used as a unit are superior to a single bottle of SPB with an equal volume of blood for the detection of bacteremia and fungemia, and that Bactec's superiority is not due to the method of detection.

Bacteria↗

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.

Bacteria↗

Clinical comparison of an agar slide blood culture bottle with tryptic soy broth and a conventional blood culture bottle with supplemented peptone broth.

The Roche Septi-Chek biphasic blood culture system with tryptic soy broth was compared with a conventional blood culture bottle with supplemented peptone broth in 6,956 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 aureus, S. epidermidis, Escherichia coli and other Enterobacteriaceae, Pseudomonas aeruginosa, and Candida albicans and C. tropicalis were recovered from the biphasic system (P less than 0.001). In contrast, more clinically important anaerobic bacteria (P less than 0.001) and Gardnerella vaginalis (P less than 0.05) were recovered in conventional supplemented peptone broth. Staphylococci (P less than 0.01), Enterobacteriaceae other than E. coli (P less than 0.05), and fungi (P less than 0.001) were detected 1 or more days earlier in the biphasic system, whereas streptococci (P less than 0.001) were detected earlier in the conventional bottle. The overall superiority of the agar slide blood culture system compared with conventional blood culture bottles was confirmed by this evaluation. For optimal detection of anaerobic bacteremia, however, the agar slide bottle should be paired with an anaerobic bottle.

Agar↗

Effect of atmosphere and duration of incubation on primary isolation of group A streptococci from throat cultures.

The optimal incubation conditions for isolation of group A streptococci from throat cultures are controversial. Therefore, we compared the effects of aerobic and anaerobic incubations after 24 and 48 h on the recovery of group A streptococci. Throat swabs submitted to the clinical laboratory were inoculated onto duplicate 5% sheep blood agar plates, incubated aerobically or anaerobically (GasPak jar) at 35 degrees C, and examined semiquantitatively after 24 and 48 h. Group A streptococci were identified by the fluorescent-antibody technique. Of 1,040 specimens, 506 (48.6%) grew beta-hemolytic streptococci, including 200 (19.2%) group A streptococci. Group A streptococci were recovered significantly more often with anaerobic incubation than with aerobic incubation after 24 h (182 versus 138; P less than 0.001) and after 48 h (193 versus 174; P less than 0.05). Non-group A beta-hemolytic streptococci also were recovered significantly more often with anaerobic incubation after 24 and 48 h (P less than 0.001). Colony counts were not affected by the incubation atmosphere. We conclude that incubation of throat cultures in an anaerobic atmosphere is superior to incubation in air for detection of group A streptococci. The greater sensitivity of anaerobic incubation, however, may not justify the extra laboratory effort and cost required to differentiate group A streptococci from the non-group A streptococci detected as a result of anaerobic incubation. Throat cultures should be examined after 24 and 48 h, especially if plates are incubated aerobically.

Adult↗

Controlled evaluation of hypertonic sucrose medium at a 1:5 ratio of blood to broth for detection of bacteremia and fungemia in supplemented peptone broth.

The value of hypertonic media in the detection of bacteremia and fungemia is controversial, since prior clinical trials have yielded conflicting results with different media. Earlier, we showed that the addition of 10% sucrose to supplemented peptone broth at a 1:10 ratio of blood to broth yielded better recovery of Staphylococcus epidermidis, the Enterobacteriaceae, Pseudomonas aeruginosa, and yeasts. To evaluate the effect of 10% sucrose on blood cultured at a 1:5 ratio, we compared the yield and speed of detection of clinically important microorganisms from adult patients in 5,839 blood samples cultured in supplemented peptone broth with 0.03% sodium polyanetholesulfonate with and without 10% sucrose. The atmosphere of incubation (open venting units), 1:5 ratio of blood to broth, and methods of processing were the same for both bottles. Recovery of facultative gram-positive (P less than 0.02) and gram-negative (P less than 0.02) bacteria was improved, but the recovery of anaerobic gram-negative bacteria was both reduced (P less than 0.01) and delayed (P less than 0.02) by sucrose. The total yield of microorganisms including fungi, however, was increased with sucrose. The effect of sucrose on blood cultures appears to depend on the ratio of blood to broth as well as on the medium used and strains of microorganisms encountered.

Bacteria↗

Dark-field microscopy of human feces for presumptive diagnosis of Campylobacter fetus subsp. jejuni enteritis.

To determine the value of direct dark-field microscopy for diagnosing enteritis due to Campylobacter fetus subsp. jejuni, we examined 1,377 human fecal specimens for bacteria with typical Campylobacter darting motility, leukocytes, and erythrocytes. Eighty-four specimens (6.1%) grew C.fetus subsp. jejuni. Of the 48 specimens showing Campylobacter motility, 30 (62%) grew C. fetus subsp. jejuni. The sensitivity, specificity, and predictive value of observing Campylobacter motility were 36%, 99%, and 62%, respectively. The predictive value of detecting Campylobacter motility was improved if the specimens were diarrheal (23 of 31, 74%), leukocytes were present (25 of 33, 76%), erythrocytes were present (22 of 27, 81%), or if all of the above findings were present (18 of 20, 90%). The sensitivity of detecting Campylobacter darting motility was highest if specimens were examined within 2 h of arrival in the laboratory (15 of 30, 50%) as opposed to after 2 h (15 of 53, 28%; P less than 0.01). Prompt dark-field microscopic examination of diarrheal stool specimens is valuable for the presumptive diagnosis of Campylobacter enteritis.

Bacteriological Techniques↗

Controlled evaluation of the volume of blood cultured in detection of bacteremia and fungemia.

To evaluate the role of the volume of blood cultured in the detection of clinically important bacteremia and fungemia in adults, we evaluated the yield and speed of detection of microorganisms from 5,317 paired 2- and 5-ml samples of blood. The same kind of medium (supplemented peptone broth with 0.03% sodium polyanetholsulfonate) and atmosphere of incubation (open venting units) were used for all blood cultures. Only adequately filled (less than or equal to 80% of stated volume) sets (20-ml tube and 50-ml bottle) were compared statistically. Significantly more bacteria (p less than 0.01), Pseudomonas spp. In particular (P less than 0.05), were isolated from the 5-ml samples of blood. We conclude that the volume of blood cultured is a critical factor in the detection of septicemia. Consequently, valid evaluation of other factors influencing the detection of septicemia must be based on comparisons in which equal volumes of blood are cultured.

Adult↗

Comparison of acridine orange, methylene blue, and Gram stains for blood cultures.

Direct microscopic screening of blood cultures by Gram stain or methylene blue stain is time consuming and frequently insensitive. Therefore, we evaluated a fluorescent-staining procedure that uses acridine orange (AO) at pH 3.5 and compared it with the methylene blue and Gram stain procedures. All smears were prepared within 24 h of receiving the culture, fixed with methanol, and examined without the results of the companion smears being known. AO-stained smears were examined with incident-light fluorescence at 600 x magnification and confirmed at 1,500x magnification. All bottles macroscopically positive within 24 h were excluded from the study. Of 2,946 cultures entered into the study, 204 (6.9%) were positive within 3 days. The sensitivity and specificity of AO based on these culture results were 52 and 98%, respectively, compared with 38% sensitivity and 99% specificity by methylene blue and Gram stains. The AO staining procedure is a simple, sensitive, screening technique for the early detection of positive blood cultures.

Acridine Orange↗