1.Direct Identification and Antimicrobial Susceptibility Testing of Bacteria From Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry and the Vitek 2 System.
Sung Jin JO ; Kang Gyun PARK ; Kyungja HAN ; Dong Jin PARK ; Yeon Joon PARK
Annals of Laboratory Medicine 2016;36(2):117-123
BACKGROUND: We evaluated the reliability and accuracy of the combined use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial identification and Vitek 2 antimicrobial susceptibility testing (AST) for bacteria from positive blood culture bottles. METHODS: Direct identification and AST were performed in parallel to the standard methods in monomicrobial positive blood culture bottles. In total, 254 isolates grown on aerobic and/or anaerobic bottles were identified with MALDI-TOF Vitek MS (bioMerieux, France), and 1,978 microorganism/antimicrobial agent combinations were assessed. For isolates from anaerobic bottles, an aliquot of the culture broth was centrifuged, washed, and filtered through a nylon mesh. For isolates from aerobic/pediatric bottles, a lysis step using 9.26% ammonium chloride solution and 2% saponin solution was included. RESULTS: The overall correct identification rate was 81.8% (208/254) and that for gram-positive/gram-negative isolates was 73.9%/92.6%, respectively, and it was 81.8%, 87.6%, and 57.9% for isolates from aerobic, anaerobic, and pediatric bottles, respectively. Identification was not possible in 45 cases, and most of these isolates were streptococci (N=14) and coagulase-negative staphylococci (N=11). Misidentification occurred only in one case. Compared with standard methods, direct AST showed 97.9% (1,936/1,978) agreement with very major error of 0.25%, major error of 0.05%, and minor error of 1.8%. CONCLUSIONS: This simple and cost-effective sample preparation method gives reliable results for the direct identification and AST of bacteria. For the identification of streptococci and coagulase-negative staphylococci, the method should be further improved.
Adult
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Ammonium Chloride/chemistry
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Anti-Infective Agents/*pharmacology
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Child
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Gram-Negative Bacteria/drug effects/*isolation & purification/metabolism
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Gram-Positive Bacteria/drug effects/*isolation & purification/metabolism
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Humans
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Reagent Kits, Diagnostic
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Saponins/chemistry
;
*Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
2.An experimental study of effect of different extracts of Alisma orientalis on urinary calcium oxalate stones formation in rats.
Zheng-guo CAO ; Ji-hong LIU ; A M RADMAN ; Ji-zhou WU ; Chun-ping YING ; Si-wei ZHOU
China Journal of Chinese Materia Medica 2003;28(11):1072-1075
OBJECTIVETo study the effect of different extracts of Alisma orientalis on urinary calcium oxalate stone formation in rats and to identify the effective constituents.
METHODDifferent extracts were administered through a stomach tube to rats of different groups with renal calcium oxalate stones induced by ethylene glycol (EG) and ammonium chloride (AC).
RESULTIn the rats administered with ethyl acetate elution of ethyl acetate extract, blood Cr, BUN, renal tissue calcium content, urinary calcium excretion and crystals deposition in renal tissue were significantly lower than those of the stone formation group.
CONCLUSIONThe ethyl acetate elution of ethyl acetate fraction extract of Alisma orientalis can significantly inhibit urinary calcium oxalate stone formation in rats and be the most effective constituent of Alisma orientalis.
Alisma ; chemistry ; Ammonium Chloride ; Animals ; Blood Urea Nitrogen ; Calcium ; metabolism ; Calcium Oxalate ; urine ; Creatinine ; blood ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ethylene Glycol ; Kidney ; metabolism ; Kidney Calculi ; chemically induced ; metabolism ; prevention & control ; Magnesium ; metabolism ; urine ; Male ; Rats ; Rats, Wistar
3.Effect of Rongshi granule on renal stone formation and osteopontin expression in rat urolithiasis model.
Ping HUANG ; Su-Wen YANG ; Wei-Hua HUANG ; Fan-Zhi KONG ; Yi-Jia LOU
China Journal of Chinese Materia Medica 2006;31(14):1188-1191
OBJECTIVETo investigate the effect of Rongshi granule on osteopontin(OPN) expression.
METHODThe urlisthiasis rats were induced by ethylene glycol (EG) and ammonium chloride, the control group rats were non-treated, and the Rongshi granule groups (low-dose group, middle-dose group and high-dose group) were administered Rongshi granule in addition to EG and ammonium chloride in 21 days. Pooled 24 h urine samples from each group were collected weekly with the use of metabolic cages, the concentration of uric calcium and oxalic acid were respectively measured by EDTA and photoelectric colorimetric method. Eight animals from each group were killed at 0, 7, 14, and 21 days, kidneys were histologic examinaed and immunohistochemical staining.
RESULTThe expression of kidney osteopontin in model group was obviously higher than that of control group (P <0.01), and was up to the highest at 21 days with 1.4 times (0.281 3/0.201 8) of the control group. The expression of kidney osteopontin in all of the Rongshi granule groups were lower than those of model group (P < 0.05), with an obvious dose-dependent manner. The degree of the kidney calcium oxalate crystal of the rats in all the Rongshi granule groups was much lower than that of model group, and the uric calcium and oxalic acid were much lower than those of model group (P < 0.01).
CONCLUSIONThe Rongshi granule could inhibit the expression of osteopontin in rat urolithiasis model.
Ammonium Chloride ; Animals ; Calcium ; urine ; Calcium Oxalate ; metabolism ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Ethylene Glycol ; Female ; Kidney ; metabolism ; Kidney Calculi ; chemically induced ; metabolism ; Male ; Osteopontin ; metabolism ; Oxalic Acid ; urine ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley