Rapid detection (4 h) of methicillin-resistant Staphylococcus aureus by a bioluminescence method

Park, C H; Hixon, Deborah L.; McLaughlin, C M; Cook, J. F.

doi:10.1128/jcm.26.6.1223-1224.1988

Cited by 9 publications

(4 citation statements)

References 8 publications

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“…Several approaches have been investigated in the past for the performance of rapid antibiotic susceptibility testing, including ATP-bioluminescence (6,8,9,15,18,19,22,29,31,32), turbidimetry (1), impedance (4), conductance (23), and radiometry (5). The ATP-bioluminescence method was notable, because several difficulties related to reagents and instrumentation had been overcome (12,13,27,28,32).…”

Section: Discussionmentioning

confidence: 99%

Novel Antibiotic Susceptibility Tests by the ATP-Bioluminescence Method Using Filamentous Cell Treatment

Hattori

Nakajima

Ohara

et al. 1998

Antimicrob Agents Chemother

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Antimicrobial susceptibility testing by the ATP-bioluminescence method has been noted for its speed; it provides susceptibility results within 2 to 5 h. However, several disagreements between the ATP method and standard methodology have been reported. The present paper describes a novel ATP method in a 3.5-h test which overcomes these deficiencies through the elimination of false-resistance discrepancies in tests on gram-negative bacteria with β-lactam agents. In our test model using Pseudomonas aeruginosa and piperacillin, it was shown that ATP in filamentous cells accounted for the false resistance. We found that 0.5% 2-amino-2-methyl-1,3-propanediol (AMPD) extracted ATP from the filamentous cells without affecting normal cells and that 0.3 U of adenosine phosphate deaminase (APDase)/ml simultaneously digested the extracted ATP. We used the mixture of these reagents for the pretreatment of cells in a procedure we named filamentous cell treatment, prior to ATP measurements. This novel ATP method with the filamentous cell treatment eliminated false-resistance discrepancies in tests on P. aeruginosa with β-lactam agents, including piperacillin, cefoperazone, aztreonam, imipenem-cilastatin, ceftazidime, and cefsulodin. Furthermore, this novel methodology produced results which agreed with those of the standard microdilution method in other tests on gram-negative and gram-positive bacteria, including P. aeruginosa, Escherichia coli,Staphylococcus aureus, and Enterococcus faecalis, for non-β-lactam agents, such as fosfomycin, ofloxacin, minocycline, and aminoglycosides. MICs obtained by the novel ATP method were also in agreement with those obtained by the agar dilution method of susceptibility testing. From these results, it was shown that the novel ATP method could be used successfully to test the activities of antimicrobial agents with the elimination of the previously reported discrepancies.

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Section: Discussionmentioning

confidence: 99%

Novel Antibiotic Susceptibility Tests by the ATP-Bioluminescence Method Using Filamentous Cell Treatment

Hattori

Nakajima

Ohara

et al. 1998

Antimicrob Agents Chemother

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“…We consider the use of a more nutritious culture media as a possible way to hasten the bacterial growth, if the use of these media does not distort the results of the MIC assessment. The approach of using rich nutrient media (Todd-Hewitt broth) to speed up the AST has been successfully used before [26]. Such an approach may work with carbapenemase-producing K. pneumoniae strains: Although faster growth would likely stimulate the effect of the antibiotic, the production of carbapenemase can also increase, which could balance the effect of the antibiotic and allow the obtaining of an early result.…”

Section: Discussionmentioning

confidence: 99%

Fluorescence Microscopy: Determination of Meropenem Activity against Klebsiella pneumoniae

et al. 2023

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The development and implementation of diagnostic methods that allow rapid assessment of antibiotic activity against pathogenic microorganisms is an important step towards antibiotic therapy optimization and increase in the likelihood of successful treatment outcome. To determine whether fluorescence microscopy with acridine orange can be used for rapid assessment (≤8 h) of the meropenem activity against Klebsiella pneumoniae, six isolates including three OXA-48-carbapenemase-producers were exposed to meropenem at different levels of its concentration (0.5 × MIC, 1 × MIC, 8 or 16 µg/mL) and the changes in the viable counts within 24 h were evaluated using fluorescence microscopy and a control culture method. The approach was to capture the regrowth of bacteria as early as possible. Within the first 8 h fluorescence microscopy allowed to categorize 5 out of 6 K. pneumoniae strains by their meropenem susceptibility (based on the MIC breakpoint of 8 mg/L), but meropenem activity against three isolates, two of which were OXA-48-producers, could not be accurately determined at 8 h. The method proposed in our study requires improvement in terms of accelerating the bacterial growth and regrowth for early meropenem MIC determination. Volume-dependent elevation in meropenem MICs against OXA-48-producers was found and this phenomenon should be studied further.

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“…There are many studies comparing the abilities of different methods to detect methicillin resistance in S. aureus. They usually employ colonies of previously identified S. aureus isolates (9,(23)(24)(25)31) that have been screened for growth on selective agar with methicillin or oxacillin (1, 2, 8, 10, 13-15, 27, 28, 30), or they employ different standardized susceptibility testing methods for the discrimination of MRSA from susceptible or borderline-resistant S. aureus (6,16,29). A few studies have evaluated the simultaneous detection and identification of MRSA isolates by plating clinical surveillance specimens in selective agar media (8,14,30,32).…”

Section: Discussionmentioning

confidence: 99%

Soft Salt-Mannitol Agar–Cloxacillin Test: a Highly Specific Bedside Screening Test for Detection of Colonization with Methicillin-Resistant Staphylococcus aureus

et al. 1998

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The early detection of colonization with methicillin-resistantStaphylococcus aureus (MRSA) of patients in intensive-care units is an essential step in the strategy for preventing MRSA epidemics. In this study, tubes containing soft salt-mannitol agar with cloxacillin (6 μg/ml) (SSMAC) were prepared for inoculation of clinical samples at patients’ bedsides by personnel of an intensive-care unit. A total of 1,914 swabs from different sample sites of 81 patients were dipped into SSMAC tubes, and after 24 h of incubation (in an incubator located near the intensive-care unit), an evident color change was considered by the intensive-care-unit personnel to be an MRSA alarm. Sixty-three (3.3%) SSMAC tubes were considered positive for MRSA, 1,827 (95.4%) were considered negative, and 24 (1.2%) were considered intermediate. Compared with values for parallel conventional surveillance cultures for MRSA, excluding tubes with intermediate results, the SSMAC test had a sensitivity of 72.7%, a specificity of 99.2%, a positive predictive value of 76.2%, and a negative predictive value of 99.0%. When intermediate tubes were considered positive, the corresponding values were 75.3, 98.2, 63.2, and 99.0%, respectively. The sensitivity and specificity values of the test to identify MRSA-colonized patients were 89.4 and 100%, respectively. Oropharyngeal and naris specimens were the most reliable samples for MRSA detection. False-negative results were frequent in bronchial aspirates with low (<103 to 106CFU/ml) MRSA counts. False-positive results were mainly due to methicillin-resistant Staphylococcus haemolyticus. The SSMAC tube is a useful, rapid, and inexpensive tool for the early identification of MRSA-colonized patients and, consequently, for the implementation of measures to prevent the spread of MRSA.

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Rapid detection (4 h) of methicillin-resistant Staphylococcus aureus by a bioluminescence method

Cited by 9 publications

References 8 publications

Novel Antibiotic Susceptibility Tests by the ATP-Bioluminescence Method Using Filamentous Cell Treatment

Novel Antibiotic Susceptibility Tests by the ATP-Bioluminescence Method Using Filamentous Cell Treatment

Fluorescence Microscopy: Determination of Meropenem Activity against Klebsiella pneumoniae

Soft Salt-Mannitol Agar–Cloxacillin Test: a Highly Specific Bedside Screening Test for Detection of Colonization with Methicillin-Resistant Staphylococcus aureus

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