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Application of Rapid Detection Method Based on NG-Test® CARBA 5 in Bloodstream Infections Associated With Carbapenem-Resistant Enterobacterales
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摘要:目的 比较快速检测方法（简称快速法）与传统检测方法（简称传统法）对血培养阳性瓶的病原体鉴定、药敏试验和碳青霉烯酶型检测的一致性与准确性。方法 收集2022年3月–2022年5月血流感染标本中涂片报告结果为“革兰阴性杆菌”的血培养阳性标本51份。采用快速法对阳性血培养标本进行快速药敏试验（rapid antibiotic susceptibility test, RAST）和鉴定，根据RAST判读标准，利用NG-Test® CARBA 5试剂盒对亚胺培南耐药菌株进行酶型快速检测，结果采用PCR确认。同时采用传统法对血培养阳性标本纯培养后的菌落进行鉴定、VITEK 2 Compact药敏分析和酶型检测。结果 细菌鉴定中，两种方法鉴定大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌和鲍曼不动杆菌的一致率均为100%。药敏试验中，两种方法的符合率较高，亚胺培南符合率为100%。碳青霉烯酶型鉴定中，传统法检测出18株产丝氨酸酶与3株产金属β-内酰胺酶的肠杆菌目细菌。快速法采用试剂盒检测出18株产KPC酶、2株产NDM酶以及1株产IMP酶的血培养标本，与PCR相比，快速法检测酶型的敏感度和特异度为100%。本实验探索的快速法对血培养阳性标本进行细菌和酶型鉴定的报告时间比传统法平均节约了1.94 d。结论 本研究建立的快速法可有效缩短血培养标本报告病原微生物和药敏试验结果的时间，联合报告胶体金酶型鉴定结果可为临床医生合理使用抗菌药物、精准抗多重耐药菌感染提供参考。Abstract:Objective To compare the consistency and accuracy of a rapid test method and a traditional test method for pathogen identification, antimicrobial susceptibility and carbapenemase type identification of positive blood culture samples.Methods A total of 51 positive blood culture samples of bloodstream infection (BSI) were collected between March 2022 and May 2022. All samples were found to be “positive for Gram-negative bacilli” according to the blood smear results. The rapid method was adopted to perform rapid antimicrobial susceptibility test (RAST) and analysis of the positive blood culture samples. According to the RAST result interpretation standards, NG-Test® CARBA 5 was used for rapid carbapenemase detection of the imipenem-resistant strains and the results were confirmed by PCR. In addition, mass spectrometry, VITEK 2 Compact drug sensitivity analysis, and carbapenemase type identification were performed with the colonies cultured with positive samples according to the traditional method.Results In the identification of bacteria, the rapid method and the traditional method had 100% consistency rate in the identification results of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. In the antimicrobial susceptibility test, the consistency rate between the results of the two methods was high and the consistency rate for results for susceptibility to imipenem was 100%. In the identification of carbapenemase type, 18 serinase-producing strains and 3 metal-β-lactamase-producing strains of Enterobacterales were detected by the traditional method. With the rapid method, 18 Klebsiella pneumoniae carbapenemase (KPC)-producing strains, 2 New Delhi metallo-betalactamase (NDM)-producing strains, and 1 imipenem enzyme (IMP)-producing strain were identified in the blood culture samples by using a testing kit. Compared with the PCR results, the sensitivity and specificity of the rapid test for determining carbapenemase types were 100%. In this study, we investigated a rapid method for bacteria and carbapenemase type identification of positive blood culture specimens and found that the turnaround time (TAT) of the rapid method was reduced by 1.94 days on average in comparison with the TAT of the traditional method.Conclusion The rapid method established in the study can effectively shorten the TAT for pathogenic microorganism identification and antimicrobial susceptibility test of blood culture samples, and the joint report of colloidal gold carbapenemase type identification results can provide a reference for clinicians to use antibiotics appropriately and accurately manage multi-drug resistant bacterial infections.
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图 1 部分碳青霉烯耐药肠杆菌目细菌的快速法药敏试验结果示意图
Figure 1. Examples of antimicrobial susceptibility test of some carbapenem-resistant Enterobacteriaceae strains by the rapid method
CZA: ceftazidime/avibactam; AK: amikacin; TZP: piperacillin/tazobactam; IMP: imipenem.
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图 3 微生物实验室鉴定流程改变示意图
Figure 3. Schematic diagram of changes in the testing process in microbiology laboratory
表 1 快速法与传统法的细菌鉴定结果比较
Table 1 Comparison of the bacterium identification results between the rapid method and the traditional method
Strain Rapid method/strain Traditional method/strain Consistency rate/% Escherichia coli 19 19 100 Klebsiella pneumoniae 17 17 100 Enterobacter aerogenes 3 4 75 Enterobacter cloacae complex 2 2 100 Acinetobacter baumannii 2 2 100 Pseudomonas aeruginosa 2 2 100 Stenotrophomonas maltophilia 2 2 100 Serratia marcescens 1 2 50 Aeromonas hydrophila 0 1 0 
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表 2 快速法与传统法药敏试验结果的符合率和错误率分析
Table 2 Analysis of the coincidence rate and the error rate of the rapid method and the traditional method in antimicrobial susceptibility test
Antimicrobial agent Strain (%) CA MIE ME VME Imipenem 40 (100) 0 0 0 Piperacillin/Tazobactam 38 (95.0) 2 (5.0) 0 0 Amikacin 37 (92.5) 2 (5.0) 1 (2.5) 0 Ceftazidime/Avibactam 35 (87.5) 4 (10.0) 1 (2.5) 0 CA: complete accordance; MIE: minor error; ME: major error; VME: very major error.
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表 3 快速法与传统法的酶型鉴定结果分析
Table 3 Analysis of carbapenemase type identification results by the rapid method and the traditional method
Strain Traditional method Rapid method PCR Number Escherichia coli Serine-carbapenemase KPC blaKPC 3 Metallo-β-lactamases (MBLs) IMP blaIMP 1 Klebsiella pneumoniae Serine-carbapenemase KPC blaKPC 15 Serratia marcescens Metallo-β-lactamases (MBLs) NDM blaNDM 2 KPC: Klebsiella pneumoniae carbapenemase; IMP: imipenem enzyme; NDM: New Delhi metallo-betalactamase.
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