Role of MALDI-TOF MS in the homology analysis of vancomycin-resistant Enterococcus faecium

doi:10.3969/j.issn.1673-8640.2026.09.011

Abstract

Abstract:

Objective To analyze the role of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS） in the homology analysis of vancomycin-resistant Enterococcus faecium （VREFm）. Methods Totally，31 non-repetitive clinical isolates of VREFm from the First Affiliated Hospital of Airforce Medical University from November 2017 to November 2024 were collected. Bacterial identification and in vitro drug susceptibility tests were performed using VITEK Compact automatic bacterial identification and drug susceptibility testing instrument. The vancomycin resistance was rechecked using disk diffusion method and microbroth dilution method. The vancomycin resistance genes were detected by fluorescence quantitative polymerase chain reaction （qPCR），and homology analysis was conducted using MALDI-TOF MS. Cluster analysis was performed using VITEK-MS SARAMIS Premium software and validated by multilocus sequence typing （MLST）. Results All the 31 VREFm isolates were resistant to vancomycin，teicoplanin，penicillin，ampicillin，levofloxacin，ciprofloxacin and erythromycin. The resistance genotypes were all vanA genes. Protein mass peak clustering analysis showed that the 31 VREFm isolates could be classified into 5 major clusters. Compared with the validation results，the 27 of 31 VREFm isolates belonged to the first cluster of MALDI-TOF MS，including 19 ST78 types，4 ST80 types and 4 ST17 types；1 isolate each in the second，third，fourth and fifth clusters，namely ST389，ST192，ST78 and ST80 types. Conclusions MALDI-TOF MS has certain limitations in the homology analysis of VREFm，the consistency with MLST results is poor，and its resolution ability is weak，unable to distinguish ST types highly related to VREFm.

Key words: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, Vancomycin-resistant Enterococcus faecium, Multilocus sequence typing, Homology analysis

CLC Number:

R446

KANG Beipei, ZHENG Tian, FU Xiaorui, YANG Peihong, HE Wenfang, ZHOU Shan, ZHOU Ke. Role of MALDI-TOF MS in the homology analysis of vancomycin-resistant Enterococcus faecium[J]. Laboratory Medicine, 2026, 41(9): 891-895.

Figures/Tables 4

References 29

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基因名称	引物序列（5'~3'）	产物长度/bp
vanA	F：GCTATTCAGCTGTACTC	783
	R：CAGCGGCCATCATACGG
vanB	F：ATGGGAAGCCGATAGTC	635
	R：GATTTCGTTCCTCGACC
vanC1	F：GGTATCAAGGAAACCTC	822
	R：CTTCCGCCATCATAGCT
vanC2/C3	F：CTCCTACGATTCTCTTG	439
	R：CGAGCAAGACCTTTAAG

基因名称	引物序列（5'~3'）	产物长度/bp
vanA	F：GCTATTCAGCTGTACTC	783
	R：CAGCGGCCATCATACGG
vanB	F：ATGGGAAGCCGATAGTC	635
	R：GATTTCGTTCCTCGACC
vanC1	F：GGTATCAAGGAAACCTC	822
	R：CTTCCGCCATCATAGCT
vanC2/C3	F：CTCCTACGATTCTCTTG	439
	R：CGAGCAAGACCTTTAAG

基因名称	引物序列（5'~3'）	产物长度/bp
adk	F：GAACCTCATTTTAATGGGG	437
	R：TGATGTTGATAGCCAGACG
atpA	F：TTCAAATGGCTCATACGG	649
	R：AGTTCACGATAAGCAACAGC
ddl	F：GAGACATTGAATATGCCTTATG	560
	R：AAAAAGAAATCGCACCG
gdh	F：GGCGCACTAAAAGATATGGT	530
	R：CCAAGATTGGGCAACTTCGTCCCA
gyd	F：CAAACTGCTTAGCTCCAATGGC	395
	R：CATTTCGTTGTCATACCAAGC
purK	F：CAGATTGGCACATTGAAAG	492
	R：TTCATTCACATATAGCCCG
pstS	F：TTGAGCCAAGTCGAAGCTGGAG	583
	R：CGTGATCACGTTCTACTTCC

基因名称	引物序列（5'~3'）	产物长度/bp
adk	F：GAACCTCATTTTAATGGGG	437
	R：TGATGTTGATAGCCAGACG
atpA	F：TTCAAATGGCTCATACGG	649
	R：AGTTCACGATAAGCAACAGC
ddl	F：GAGACATTGAATATGCCTTATG	560
	R：AAAAAGAAATCGCACCG
gdh	F：GGCGCACTAAAAGATATGGT	530
	R：CCAAGATTGGGCAACTTCGTCCCA
gyd	F：CAAACTGCTTAGCTCCAATGGC	395
	R：CATTTCGTTGTCATACCAAGC
purK	F：CAGATTGGCACATTGAAAG	492
	R：TTCATTCACATATAGCCCG
pstS	F：TTGAGCCAAGTCGAAGCTGGAG	583
	R：CGTGATCACGTTCTACTTCC

抗菌药物	VSEFm			VREFm
抗菌药物	耐药	中介	敏感	耐药	中介	敏感
青霉素	94.2	0	5.8	100.0	0	0
氨苄西林	93.1	0	6.9	100.0	0	0
环丙沙星	93.6	2.3	4.1	100.0	0	0
左氧氟沙星	92.5	2.6	4.9	100.0	0	0
红霉素	91.4	5.8	2.8	100.0	0	0
利奈唑胺	0.1	0	99.9	0	0	100.0
万古霉素	0	0	100.0	100.0	0	0
替考拉宁	0	0	100.0	100.0	0	0
四环素	54.5	0.2	45.3	81.8	0	18.2
替加环素	0	0	100.0	0	0	100.0