改良Carba Np试验和mCIM/eCIM快速鉴定产碳青霉烯酶肠杆菌科细菌表型的临床应用

doi:10.3969/j.issn.1673-8640.2022.010.011

摘要/Abstract

摘要：

目的评估改良Carba Np试验与碳青霉烯灭活试验（mCIM）/乙二胺四乙酸碳青霉烯灭活试验（eCIM）在检测肠杆菌科细菌碳青霉烯酶表型中的应用价值。方法收集136株肠杆菌科细菌临床分离株，其中耐碳青霉烯类肠杆菌科细菌（CRE）86株，碳青霉烯敏感株50株。分别采用改良Carba Np试验、mCIM/eCIM进行检测，并进行表型分析和分类，以聚合酶链反应（PCR）碳青霉烯酶基因鉴定结果为标准，评价2种改良方法检测结果的差异。结果 86株CER中，耐药基因阳性81株；50株敏感菌株均未检出耐药基因。改良Carba NP试验阳性82株，mCIM/eCIM阳性78株。以PCR结果为标准，改良Carba NP试验和mCIM/eCIM检测肠杆菌科细菌碳青霉烯酶的敏感性、特异性分别为96.3%、92.7%和92.6%、94.5%，Kappa值分别为0.935和0.917。改良Carba NP试验丝氨酸碳青霉烯酶检出率为97.7%，金属碳青霉烯酶检出率为100.0%。mCIM/eCIM检测金属碳青霉烯酶的敏感性为91.2%、特异性为90.4%。改良Carba NP试验与mCIM/eCIM碳青霉烯酶阳性检出率差异无统计学意义（χ²=1.65，P>0.05）。结论改良Carba NP试验可快速、准确地鉴定CRE表型；mCIM/eCIM操作简便、干扰因素少。2种改良方法可以优势互补，有较大的临床应用价值。

关键词: 碳青霉烯耐药肠杆菌科细菌, 改良Carba Np试验, 改良碳青霉烯灭活试验

Abstract:

Objective To evaluate the application value of modified Carba Np test and modified carbapenem inactivation method（mCIM），ethylenediaminetetraacetic acid-carbapenem inactivation method（eCIM） for detecting carbapenemase producing Enterobacteriaceae phenotypes. Methods Totally，86 isolates of carbapenem resistant Enterobacteriaceae（CRE） and 50 sensitive isolates were collected. Phenotype analysis and classification were performed by modified Carba Np test and mCIM/eCIM，respectively. The carbapenemase genes were detected by polymerase chain reaction（PCR）. The difference between the 2 methods was evaluated. Results Among the 86 isolates of CRE，81 isolates were drug-resistant gene positive，and all the 50 sensitive isolates were drug-resistant gene negative. The 82 isolates of CRE were positive by modified Carba Np test，and 78 isolates were positive by mCIM/eCIM test. Compared with PCR results，the sensitivity and specificity of modified Carba Np test were 96.3% and 92.7%，respectively，the Kappa value was 0.935. The sensitivity and specificity of mCIM/eCIM test were 92.6% and 94.5%，respectively，and the Kappa value was 0.917. The detection rates of serine carbapenemase and metallocarbapenemase were 97.7% and 100.0% with modified Carba Np test. The sensitivity and specificity of mCIM/eCIM test were 91.2% and 90.4%，respectively. There was no statistical significance between modified Carba Np test and mCIM/eCIM test（χ²=1.65，P>0.05）. Conclusions The modified Carba Np test could rapidly identify the phenotypes of carbapenemase producing Enterobacteriaceae，and the modified mCIM/eCIM test is convenient，easy to operate and has few interference factors. The 2 methods could complement each other with clinical application value.

Key words: Carbapenem resistant Enterobacteriaceae, Modified Carba Np test, Modified carbapenem inactivation method

中图分类号:

R446.5

包海林, 花鸿燕, 孙恒亮, 刘华, 吉顺年, 秦陈浩, 杜鸿. 改良Carba Np试验和mCIM/eCIM快速鉴定产碳青霉烯酶肠杆菌科细菌表型的临床应用[J]. 检验医学, 2022, 37(10): 963-968.

BAO Hailin, HUA Hongyan, SUN Hengliang, LIU Hua, JI Shunnian, QIN Chenhao, DU Hong. Rapid identification of carbapenemase producing Enterobacteriaceae phenotypes by modified Carba Np test and mCIM/eCIM test[J]. Laboratory Medicine, 2022, 37(10): 963-968.

图/表 11

参考文献 17

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编号	亚胺培南西司他丁/mg	他唑巴坦/mg	EDTA/μL	（酚红+ ZnSO₄· 7H₂O）/μL	去离子水/μL
Ⅰ	0	0	0	100	900
Ⅱ	4	0	0	100	900
Ⅲ	4	4	0	100	900
Ⅳ	4	0	100	100	800
Ⅴ	4	4	100	100	800

编号	亚胺培南西司他丁/mg	他唑巴坦/mg	EDTA/μL	（酚红+ ZnSO₄· 7H₂O）/μL	去离子水/μL
Ⅰ	0	0	0	100	900
Ⅱ	4	0	0	100	900
Ⅲ	4	4	0	100	900
Ⅳ	4	0	100	100	800
Ⅴ	4	4	100	100	800

目的基因	正向引物序列（5'~3'）	反向引物序列（5'~3'）	退火温度/℃	产物大小/bp
bla_kpc-2	GTATCGCCGTCTAGTTCTGC	GGTCGTGTTTCCCTTTAGCC	56	638
bla_NDM-1	GGTTTGGCGATCTGGTTTTC	CGGAATGGCTCATCACGATC	55	621
bla_IMP-4	CTACCGCAGCAGAGTCTTTG	AACCAGTTTTGCTTACCAT	52	587
bla_VIM-1	AGTGGTGAGTATCCGACAG	ATGAAAGTGCGTGGAGAC	55	260
bla_OXA-48	GCTTGATCGCCCTCGATT	GATTTGCTCCGTGGCCGAAA	58	281

目的基因	正向引物序列（5'~3'）	反向引物序列（5'~3'）	退火温度/℃	产物大小/bp
bla_kpc-2	GTATCGCCGTCTAGTTCTGC	GGTCGTGTTTCCCTTTAGCC	56	638
bla_NDM-1	GGTTTGGCGATCTGGTTTTC	CGGAATGGCTCATCACGATC	55	621
bla_IMP-4	CTACCGCAGCAGAGTCTTTG	AACCAGTTTTGCTTACCAT	52	587
bla_VIM-1	AGTGGTGAGTATCCGACAG	ATGAAAGTGCGTGGAGAC	55	260
bla_OXA-48	GCTTGATCGCCCTCGATT	GATTTGCTCCGTGGCCGAAA	58	281

改良Carba NP 试验	mCIM	eCIM	耐药基因	耐药菌名称	菌株数/株	MIC/（μg/L）
改良Carba NP 试验	mCIM	eCIM	耐药基因	耐药菌名称	菌株数/株	美罗培南	亚胺培南
+	+	-	bla_kpc-2	肺炎克雷伯菌	36	>16	>16
+	+	+	bla_kpc-2	肺炎克雷伯菌	2	>16	>16
-	+	+	bla_kpc-2	肺炎克雷伯菌	1	>16	>16
+	-	无效	bla_kpc-2	大肠埃希菌	2	4	4
+	+	-	bla_kpc-2	阴沟肠杆菌	2	>16	>16
+	+	+	bla_NDM-1	肺炎克雷伯菌	7	>16	>16
+	+	+	bla_NDM-1	肺炎克雷伯菌	2	4	>16
+	+	+	bla_NDM-1	大肠埃希菌	16	>16	>16
+	-	无效	bla_NDM-1	大肠埃希菌	2	4	4
+	+	+	bla_NDM-1	阴沟肠杆菌	3	>16	>16
+	-	无效	bla_NDM-1	弗劳地枸橼酸杆菌	2	>16	>16
+	+	+	bla_IMP-4	肺炎克雷伯菌	3	>16	>16
+	+	-	bla_IMP-4	阴沟肠杆菌	1	>16	>16
-	+	-	bla_kpc-2+bla_NDM-1	肺炎克雷伯菌	2	>16	>16
+	+	+	未检出	肺炎克雷伯菌	2	>16	>16
-	+	-	未检出	肺炎克雷伯菌	1	>16	>16
+	-	无效	未检出	大肠埃希菌	2	>16	>16
-	-	无效	未检出	肺炎克雷伯菌	41	≤0.25	≤1.0
-	-	无效	未检出	大肠埃希菌	7	≤0.25	≤1.0
-	-	无效	未检出	阴沟肠杆菌	2	≤0.25	≤1.0