预培养对血液常见感染菌血培养阳性报警时间的影响
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摘要
目的探究不同预培养条件对血液常见感染菌血培养阳性报警时间的影响,指导临床血培养标本合理运输与接收。方法将金黄色葡萄球菌、大肠埃希菌和铜绿假单胞菌三种血液常见感染菌配成10CFU/ml和100CFU/ml浓度菌液,分别注入标准需氧和厌氧血培养瓶,人工模拟构建阳性血流感染标本。血培养瓶置于25℃和37℃预培养0、2、4、8、12、24h立即放入BacT/Alert 3D血培养仪,记录、分析阳性报警时间。结果金黄色葡萄球菌、大肠杆菌在25℃预培养8h、37℃预培养4h阳性报警时间均明显缩短,铜绿假单胞菌在25℃预培养12h、37℃预培养8h阳性报警时间明显缩短,与未预培养组(0h)相比差异均有统计学意义(P<0.05)。不同菌落数(菌液浓度)和不同培养瓶类型(需氧或厌氧)间阳性报警时间差异不明显(P>0.05)。铜绿假单胞菌25℃和37℃预培养24hBacT/Alert 3D血培养仪均报告阴性(假阴性)。结论从实验结果得出,三种实验菌的阳性报警时间主要受预培养温度和时间的影响,与注入菌落数及培养瓶类型相关性不大;不能及时放入血培养系统的标本应置于37℃预培养,预培养时间不能超过24h。
Objective To explore the effect of preincubation conditions on the time to detection(TTD)of blood culture of common blood infectious bacteria and provide evidence for shipping and handling the clinical blood culture specimens reasonably. Methods In order to mimic blood infection specimens,Staphylococcus aureus(S.aureus),Escherichia coli(E.coli)and Pseudomonas aeruginosa(P.aeruginosa)at 10 and 100CFU/ml were inoculated into standard aerobic or anaerobic blood culture bottles,and then were preincubated at 25℃ or 37℃for 0,2,4,8,12 or 24h.TTD was monitored using BacT/Alert 3Dsystem. Results Compared with the groups without preincubation,TTD was significantly shortened following preincubation for 8hr at 25℃and 4hr at 37℃for S.aureus and E.coli,as well as for 12 hr at 25℃and 8hr at37℃ for P.aeruginosa(P<0.05).No statistical differences in TTD were observed between different bacterial concentrations(P>0.05),or between aerobic and anaerobic incubation groups(P>0.05).The BacT/Alert 3Dsystem reported false-negative results for P.aeruginosa following preincubation for 24 hr at 25℃ or 37℃. Conclusions TTD of Staphylococcus aureus,Escherichia coli and Pseudomonas aeruginosain blood specimens is mainly affected by temperature and duration of preincubation rather than by input amount of CFU or type of blood culture bottle.So blood specimens that can't be put in BacT/Alert 3Dsystem in time should be preincubated at 37℃,but for no more than 24 h.
Objective To explore the effect of preincubation conditions on the time to detection(TTD)of blood culture of common blood infectious bacteria and provide evidence for shipping and handling the clinical blood culture specimens reasonably. Methods In order to mimic blood infection specimens,Staphylococcus aureus(S.aureus),Escherichia coli(E.coli)and Pseudomonas aeruginosa(P.aeruginosa)at 10 and 100CFU/ml were inoculated into standard aerobic or anaerobic blood culture bottles,and then were preincubated at 25℃ or 37℃for 0,2,4,8,12 or 24h.TTD was monitored using BacT/Alert 3Dsystem. Results Compared with the groups without preincubation,TTD was significantly shortened following preincubation for 8hr at 25℃and 4hr at 37℃for S.aureus and E.coli,as well as for 12 hr at 25℃and 8hr at37℃ for P.aeruginosa(P<0.05).No statistical differences in TTD were observed between different bacterial concentrations(P>0.05),or between aerobic and anaerobic incubation groups(P>0.05).The BacT/Alert 3Dsystem reported false-negative results for P.aeruginosa following preincubation for 24 hr at 25℃ or 37℃. Conclusions TTD of Staphylococcus aureus,Escherichia coli and Pseudomonas aeruginosain blood specimens is mainly affected by temperature and duration of preincubation rather than by input amount of CFU or type of blood culture bottle.So blood specimens that can't be put in BacT/Alert 3Dsystem in time should be preincubated at 37℃,but for no more than 24 h.
引文
[1]方丹,陈建辉.2006-2011年某医院血培养病原菌分布及耐药性回顾性分析[J].实用预防医学,2012,19(11):1726-1728.
[2]Clinical and Laboratory Standards Institute.Principles and procedures for blood cultures;approved guidline.CLSI document M47-A.Wayne,PA:Clinical and Laboratory Standards Institute,2007.
[3]Saito T,Iinuma Y,Takakura S,et al.Delayed insertion of blood culture bottles into automated continuously monitoring blood culture systems increases the time from blood sample collection to the detection of microorganisms in bacteremic patients[J].J Infect Chemother,2009,15:49-53.
[4]黄磊,孙立颖,严岩.血培养阳性报警时间在大肠埃希菌血流感染患者预后评估中的应用价值研究[J].中国全科医学,2014,17(2):162-165.
[5]Janapatla RP,Yan JJ,Chien ML,et al.Effect of overnight storage of blood culture bottles on bacterial detection time in the BACTEC9240blood culture system[J].J Microbiol Immunol Infec,2010,43(2):126-132.
[6]王沛,何永贵,陈学兵.阳性报警时间法快速鉴定血培养中的葡萄球菌[J].检验医学,2009,11:850-851.
[7]毛小琴,周光,赵梅.血培养仪阳性报警标本的多参数分析[J].中华医院感染学杂志,2014,24(22):5519-5521.
[8]Reimer LG,Wilson ML,Weinstein MP.Update on detection of bacteremia and fungemia[J].Clin Microbiol Rev,1997,10:444-465.
[9]Lee DH,Koh EH,Choi SR,et al.Growth dynamics of Staphylococcus aureus,Escherichia coli,and Pseudomonas aeruginosa as a function of time to detection in BacT/Alert 3Dblood culture bottles with various preincubation conditions[J].Ann Lab Med,2013,33:406-409.
[10]ViganòEF,Vasconi E,Agrappi C,et al.Use of simulated blood cultures for time to detection comparison between BacT/ALERT and BACTEC 9240blood culture systems[J].Diagn Microbiol Infect Dis,2002,44:235-240.
[11]van der Velden LB,Vos FJ,Mouton JW,et al.Clinical impact of preincubation of blood cultures at 37℃[J].J Clin Microbiol,2011,49:275-280.
[12]Kerremans JJ,van der Bij AK,Goessens W,et al.Immediate incubation of blood cultures outside routine laboratory hours of operation accelerates antibiotic switching[J].J Clin Microbiol,2009,47:3520-3523.
[13]Sautter RL,Bills AR,Lang DL,et al.Effects of delayed-entry conditions on the recovery and detection of microorganisms from BacT/ALERT and BACTEC blood culture bottles[J].J Clin Microbiol,2006,44:1245-1249.
[2]Clinical and Laboratory Standards Institute.Principles and procedures for blood cultures;approved guidline.CLSI document M47-A.Wayne,PA:Clinical and Laboratory Standards Institute,2007.
[3]Saito T,Iinuma Y,Takakura S,et al.Delayed insertion of blood culture bottles into automated continuously monitoring blood culture systems increases the time from blood sample collection to the detection of microorganisms in bacteremic patients[J].J Infect Chemother,2009,15:49-53.
[4]黄磊,孙立颖,严岩.血培养阳性报警时间在大肠埃希菌血流感染患者预后评估中的应用价值研究[J].中国全科医学,2014,17(2):162-165.
[5]Janapatla RP,Yan JJ,Chien ML,et al.Effect of overnight storage of blood culture bottles on bacterial detection time in the BACTEC9240blood culture system[J].J Microbiol Immunol Infec,2010,43(2):126-132.
[6]王沛,何永贵,陈学兵.阳性报警时间法快速鉴定血培养中的葡萄球菌[J].检验医学,2009,11:850-851.
[7]毛小琴,周光,赵梅.血培养仪阳性报警标本的多参数分析[J].中华医院感染学杂志,2014,24(22):5519-5521.
[8]Reimer LG,Wilson ML,Weinstein MP.Update on detection of bacteremia and fungemia[J].Clin Microbiol Rev,1997,10:444-465.
[9]Lee DH,Koh EH,Choi SR,et al.Growth dynamics of Staphylococcus aureus,Escherichia coli,and Pseudomonas aeruginosa as a function of time to detection in BacT/Alert 3Dblood culture bottles with various preincubation conditions[J].Ann Lab Med,2013,33:406-409.
[10]ViganòEF,Vasconi E,Agrappi C,et al.Use of simulated blood cultures for time to detection comparison between BacT/ALERT and BACTEC 9240blood culture systems[J].Diagn Microbiol Infect Dis,2002,44:235-240.
[11]van der Velden LB,Vos FJ,Mouton JW,et al.Clinical impact of preincubation of blood cultures at 37℃[J].J Clin Microbiol,2011,49:275-280.
[12]Kerremans JJ,van der Bij AK,Goessens W,et al.Immediate incubation of blood cultures outside routine laboratory hours of operation accelerates antibiotic switching[J].J Clin Microbiol,2009,47:3520-3523.
[13]Sautter RL,Bills AR,Lang DL,et al.Effects of delayed-entry conditions on the recovery and detection of microorganisms from BacT/ALERT and BACTEC blood culture bottles[J].J Clin Microbiol,2006,44:1245-1249.