HB&L微生物培养系统用于脑脊液快速培养鉴定及药敏试验的方法研究

HB&L microbial culture system for rapid identification and drug susceptibility testing of cerebrospinal fluid

  • 摘要:
      背景  脑脊液标本病原菌培养是颅内感染诊断的金标准,目前临床多采用血培养瓶对脑脊液标本进行培养,阳性报警后再经过传代培养方可进行菌种鉴定和药敏试验,其耗时长,不利于临床及时诊治,有必要改进方法提高报告时效。
      目的  探讨HB&L(人体生物体液)微生物培养系统快速培养脑脊液阳性标本后进行菌种鉴定和药敏试验的准确性和时效性。
      方法  收集2021年1月- 2022年1月我中心血液培养仪报阳性的脑脊液样本37例,分别进行常规分离培养和HB&L快速培养,以激光解析电离飞行时间质谱仪(MALDI-TOF MS)进行鉴定,以VITEK-2 Compact细菌鉴定药敏仪进行药敏试验。以常规培养法为金标准,评价HB&L快速培养法鉴定结果的正确率和药敏结果的符合率,比较两种方法的周转时间。
      结果  共分离鉴定出37株菌株,其中11株凝固酶阴性葡萄球菌、1株金黄色葡萄球菌、1株粪肠球菌、2株纹带棒杆菌、1株蜡样芽孢杆菌、6株鲍曼不动杆菌、6株肺炎克雷伯菌、3株铜绿假单胞菌、2株大肠埃希菌、1株阴沟肠杆菌、1株嗜麦芽窄食单胞菌和2株真菌。HB&L快速培养法菌种鉴定正确率为100%。与常规法药敏试验结果比较,HB&L快速培养法鉴定革兰阴性菌时替加环素、阿米卡星和米诺环素药物符合率为94.44%,其他抗菌药物符合率为100%;革兰阳性菌,红霉素符合率为92.31%,其他抗菌药物符合率为100%;HB&L快速培养法标准总符合率为98.90%,一般错误为0.82%,严重错误为0,极严重错误为0.27%。HB&L快速培养法菌种鉴定和药敏试验的周转时间相比常规法分别缩短22.10 h和24.77 h,差异有统计学意义(P<0.001)。
      结论  HB&L联合MALDI-TOF MS和VITEK-2 Compact可用于脑脊液培养阳性样本进行快速鉴定和药敏试验,回报时间缩短24 h,为临床及时治疗提供依据。

     

    Abstract:
      Background   Detecting pathogen from the culturing cerebrospinal fluid (CSF) is the golden standard for the diagnosis of intracranial infection. At present, blood culture bottles are commonly used to culture CSF samples, and subculture is adopted for positive ones before pathogen identification and antimicrobial susceptibility testing, which is time-consuming and unavailable for timely diagnosis and treatment. So it is necessary to increase the report efficient by refining the process.
      Objective  To explore the accuracy and efficiency of the species identification and antimicrobial susceptibility testing of positive samples after rapidly culturing by HB&L (Human biological fluid) microbial culture system.
      Methods  A total of 37 CSF positive samples were collected in the Sixth Medical Center of Chinese PLA General Hospital from January 2021 to January 2022. These samples were processed with the conventional isolation culture method and HB&L rapid culture method, respectively. Then the species identification and antimicrobial susceptibility test were performed with MALDI-TOF MS and VITEK-2 Compact system. The conventional culture method was defined as golden standard and the accuracy of identification and coincidence rate of antimicrobial susceptibility testing were assessed, as well as the turnaround time of the two methods.
      Results  Totally 37 strains were isolated and identified, including coagulase-negative Staphylococci (11 strains), S.aureus (1 strains), E.faecali s (1 strains), Corynebacterium striatum (2 strains), Bacillus cereus (1 strains), A.baumannii (6 strains), K.pneumoniae (6 strains), P.aeruginosa (3 strains), E.coli (2 strains), E.cloacae (1 strains), S.maltophilia (1 strains) and fungus (2 strains). The identification accuracy of the HB&L rapid culture method was 100.00%. Compared with the conventional culture method, the antimicrobial susceptibility test coincidence rate of HB&L rapid culture method for Gram-negative bacteria was 100.00%, except for tegacyclin, amikacin and minocycline with the coincidence rate of 94.44%. The antimicrobial susceptibility test coincidence rate of Gram-positive bacteria was 100.00%, except for erythromycin with the coincidence rate of 92.31%. The total coincidence rate of the antimicrobial susceptibility test was 98.90%, with MIE of 0.82%, ME of 0.00% and VME of 0.27%. The turnaround time of species identification and antimicrobial susceptibility test of HB&L rapid culture method were 22.10 h and 24.77 h, which were shorter than those of conventional method (P<0.001, respectively).
      Conclusion  The HB&L system, combining with MALDI-TOF MS and VITEK-2 Compact system, can be used in species identification and antimicrobial susceptibility testing of the positive CSF samples, which can provide evidences for clinical timely treatment by shortening the report time.

     

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