胶体金免疫层析技术与mCIM联合eCIM在细菌碳青霉烯酶检测中的效能与应用评估

Application and evaluation of colloidal gold immunochromatography and modified carbapenem inactivation method combined with EDTA-carbapenem inactivation method in detection of bacterial carbapenemase

  • 摘要:
    目的 比较胶体金免疫层析技术与改良碳青霉烯灭活试验(mCIM)联合乙二胺四乙酸改良碳青霉烯灭活试验(eCIM)两种方法在临床常见革兰阴性菌碳青霉烯酶检测中的效能,评估其临床应用价值。
    方法 153 株对碳青霉烯类药物不敏感的革兰阴性菌,均进行mCIM联合eCIM、胶体金免疫层析技术筛选。 以基因组高通量测序结果为金标准,比较两种方法检测碳青霉烯类药物不敏感的革兰阴性菌的碳青霉烯酶的灵敏度、特异度、检测时长以及对碳青霉烯酶型别的鉴定能力。
    结果 153株碳青霉烯类药物不敏感的革兰阴性菌中,基因组高通量测序和耐药基因筛查结果显示,121株携带碳青霉烯酶基因,32株不携带碳青霉烯酶基因。 mCIM联合eCIM试验检测肠杆菌目细菌碳青霉烯酶的灵敏度为92.00%,特异度为100.00%,阳性预测值为100.00%,阴性预测值为84.21%;检测鲍曼不动杆菌中碳青霉烯酶的灵敏度为73.91%,特异度为100.00%,阳性预测值为100.00%,阴性预测值为84.21%;检测铜绿假单胞菌中碳青霉烯酶的灵敏度为95.65%,特异度为100.00%,阳性预测值为100.00%,阴性预测值为96.97%。 与胶体金免疫层析技术相比,mCIM联合eCIM检测铜绿假单胞菌中碳青霉烯酶的符合率差异无统计学意义(Fisher精确检验,P=0.999);但这两种方法检测肠杆菌目细菌(Fisher精确检验,P=0.029)和鲍曼不动杆菌(Fisher精确检验,P=0.024)中碳青霉烯酶的符合率差异有统计学意义。 胶体金免疫层析技术优于mCIM联合eCIM试验。 此外,胶体金免疫层析技术的检测时长短于mCIM联合eCIM试验,且在鉴定细菌碳青霉烯酶型别的能力上优于mCIM联合eCIM试验,但测试成本较mCIM联合eCIM高。
    结论 在铜绿假单胞菌中碳青霉烯酶的检测中,胶体金免疫层析与mCIM联合eCIM试验无明显差异;在肠杆菌目细菌和鲍曼不动杆菌中碳青霉烯酶的检测中,胶体金免疫层析法灵敏度优于mCIM联合eCIM试验。 不同层级的微生物实验室可根据当地细菌中碳青霉烯酶酶型特征和患者具体状况选择合适的碳青霉烯酶检测方法。

     

    Abstract:
    Objective To evaluate the clinical application of colloidal gold immunochromatography and modified carbapenem inactivation method (mCIM) combined with Ethylene Diamine Tetraacetic Acid (EDTA) carbapenem inactivation method (eCIM) in the detection of carbapenemase in common gram-negative bacteria.
    Methods A total of 153 strains of carbapenem insensitive gram-negative bacteria were screened by mCIM combined with eCIM and colloidal gold immunochromatography. Using high-throughput sequencing results as the gold standard, the sensitivity, specificity, detection time and carbapenemase type identification power of the two methods were compared.
    Results In 153 gram-negative bacteria which were insensitive to carbapenems, 121 carried carbapenemase gene indicated by high-throughput sequencing and drug resistance gene screening. In mCIM combined with eCIM, the sensitivity, specificity, positive prediction value and negative predictive value were 92.00%, 100.00%, 100.00% and 84.21%, respectively, in the detection of carbapenemase in Enterobacteriaceae, 73.91%, 100.00%, 100.00% and 84.21%, respectively, in the detection of carbapenemase in Acinetobacter baumannii and 95.65%, 100.00%, 100.00% and 96.97%, respectively, in the detection of carbapenemase in Pseudomonas aeruginosa. Compared with colloidal gold immunochromatography, mCIM combined with eCIM showed no significant differences in the detection of carbapenemase in P. aeruginosa (Fisher's exact test, P=0.999), but showed significant differences in the coincidence rate of carbapenemase detection in Enterobacteriaceae ( Fisher's exact test, P=0.029) and A. baumannii (Fisher's exact test, P=0.024). The detection time of colloidal gold immunochromatography was shorter than that of mCIM combined with eCIM, and the performance in carbapenemase type identification was better than mCIM combined with eCIM, but the cost was higher compared with mCIM combined with eCIM.
    Conclusion There was no significant difference in the detection of carbapenemase in P. aeruginosa between the two methods. In the detection of carbapenemase in Enterobacteriaceae and A. baumannii, colloidal gold immunochromatography was more sensitive. Microbiology laboratories at different levels can select carbapenemase detection method according to the local carbapenemase type characteristics and specific conditions of patients.

     

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