16S rRNA基因序列、生化鉴定、质谱3类方法鉴定常见微生物的结果分析
|
摘要
目的比较16SrRNA基因序列、生化鉴定、质谱鉴定3类实验方法分析沙门氏菌、金黄色葡萄球菌、蜡样芽孢杆菌的鉴定结果的异同。方法挑选沙门氏菌、金黄色葡萄球菌、蜡样芽孢杆菌各50株,分别进行16S rRNA基因序列测定、VITEK COMPACT 2生化鉴定、质谱鉴定3类实验,并比较3类实验的鉴定结果。结果 3类实验方法对大部分沙门氏菌鉴定在属水平,生化鉴定方法对少数血清型鉴定到种水平;对金黄色葡萄球菌均鉴定到种水平; 16SrRNA基因序列、生化方法对蜡样芽孢杆菌鉴定到属水平,质谱鉴定到种水平。结论 3类实验方法对大部分沙门氏菌、所有金黄色葡萄球菌鉴定水平相同,质谱鉴定蜡样芽孢杆菌的结果更准确,且质谱鉴定时效性更高。
Objective To compare the differences and similarities of results of 3 types of experimental methods,including 16 S rRNA gene sequence,VITEK COMPACT 2 biochemical reaction and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) on the determination of Salmonella,Staphylococcus aureus and Bacillus cereus.Methods Fifty strains of Salmonella,Staphylococcus aureus and Bacillus cereus were selected respectively to be identified by 16 S rRNA gene sequence,VITEK COMPACT 2 biochemical identification and MALDI-TOF MS,and the identification results of the 3 kinds of experiments were compared.Results Three experimental methods were used to identify most Salmonella at the genus level.And VITEK COMPACT 2 biochemical reaction identified a few serotypes at the species level.The results of Staphylococcus aureus with the above methods were all on species.Bacillus cereus can be on species level with MALDI-TOF MS,but only on genus with the other methods.Conclusion The identification results of most Salmonella and all Staphylococcus aureus are consistent by the 3 methods,but MALDI-TOF MS can identify the Bacillus cereus accurately.MALDI-TOF MS presents higher timeliness on pathogen identification.
Objective To compare the differences and similarities of results of 3 types of experimental methods,including 16 S rRNA gene sequence,VITEK COMPACT 2 biochemical reaction and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) on the determination of Salmonella,Staphylococcus aureus and Bacillus cereus.Methods Fifty strains of Salmonella,Staphylococcus aureus and Bacillus cereus were selected respectively to be identified by 16 S rRNA gene sequence,VITEK COMPACT 2 biochemical identification and MALDI-TOF MS,and the identification results of the 3 kinds of experiments were compared.Results Three experimental methods were used to identify most Salmonella at the genus level.And VITEK COMPACT 2 biochemical reaction identified a few serotypes at the species level.The results of Staphylococcus aureus with the above methods were all on species.Bacillus cereus can be on species level with MALDI-TOF MS,but only on genus with the other methods.Conclusion The identification results of most Salmonella and all Staphylococcus aureus are consistent by the 3 methods,but MALDI-TOF MS can identify the Bacillus cereus accurately.MALDI-TOF MS presents higher timeliness on pathogen identification.
引文
[1]沈萍,陈向东.微生物学实验(第4版)[M].北京:高等教育出版社,2010.Shen P,Chen XD.Microbiology experiment(Forth Edition)[M].Beijing:Higher Education Press,2010.
[2]Dieckmann R,Malorny B.Rapid screening of epidemiologically important Salmonella enterica subsp.enterica serovars by whole-cell matrix-assisted laser desorption ionization-time of flight mass spectrometry[J].Appl Environ Microbiol,2011,77(12):4136-4146.
[3]Huang CH,Huang L.Rapid species-and subspecies-specific level classification and identification of Lactobacillus casei group members using MALDI biotyper combined with clinprotools[J].J Dairy Sci,2018,101(2):979-991.
[4]Lawton SJ,Weis AM,Byrne BA,et al.Comparative analysis of campylobacter isolates from wild birds and chickens using MALDI-TOFMS,biochemical testing,and DNA sequencing[J].J Veter Diagnost Invest,2018,30(3):354-361.
[5]Pranada AB,Witt E,Bienia M,et al.Accurate differentiation of mycobacterium chimaera from mycobacterium intracellulare by MALDI-TOF MS analysis[J].J Med Microbiol,2017,66(5):670-677.
[6]Schlebusch S,Pric GR,Gallagher RL,et al.MALDI-TOF MS meets WGS in a VRE outbreak investigation[J].Eur J Clin Microbiol Infect Dis,2017,36(3):495-499.
[7]Ayeni FA,Andersen C,Lauritsen NN.Comparison of growth on mannitol salt agar,matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,VITEK?2 with partial sequencing of 16S rRNA gene for identification of coagulase-negative staphylococci[J].Microb Pathogen,2017,105:255-259.
[8]李修远,李亚楠,鲁辛辛.MALDI-TOF MS用于临床微生物检验中常见菌鉴定的应用探讨[J].临床检验杂志,2018,46(4):297-303.Li XY,Li YN,Lu XX.Discussion on the application of MALDI-TOF MSin the identification of common bacteria in clinical microbiological examination[J].J Clin Lab Sci,2018,46(4):297-303.
[9]佚名.MALDI-TOF MS病原体鉴定质量保证专家共识[J].中华检验医学杂志,2018,41(8):567-570.Anonymous.Expert consensus on MALDI-TOF MS pathogen identification quality assurance[J].Chin J Lab Med,2018,41(8):567-570.
[10]张雪英,叶红,李爱云,等.采用MALDI-TOF MS和全基因组测序鉴定和分析动弯杆菌属及其亲缘关系[J].中华微生物学和免疫学杂志,2018,38(6):446-450.Zhang XY,Ye H,Li AY,et al.Idention and analsis of genus acinetobacter and its relatives by MALDI-TOF MS and whole genome sequencing[J].Chin J Microbiol Immunol,2018,38(6):446-450.
[11]俞静,陈峰,李媛睿,等.应用MALDI-TOF MS技术快速鉴定布鲁菌[J].国际检验医学,2018,39(12):1443-1447.Yu J,Chen F,Li YR,et al.Rapidly identifition of Brucella by MALDI-TOF MS[J].Int Lab Med,2018,39(12):1443-1447.
[12]李颖,徐英春.MALDI-TOF MS在病原性丝状真菌鉴定中的应用进展[J].中国真菌学,2017,12(6):378-381.Li Y,Xu YC.Application progress of MALDI-TOF MS in identification of pathogenic filamentous fungi[J].Chin Mycol,2017,12(6):378-381.
[13]Li Y.Christiaens GCMLP,Gille JJP,et al.Non-invasive prenatal detection of achondroplasia in size-fractionated cell-free DNA by MALDI-TOF MSassay[J].Prenat Diagn,2007,27(1):11-17.
[14]Piseth Seng CA,Rolain JM,et al.Identification of rare pathogenic bacteria in a clinical microbiologylaboratory:impact of matrix-assisted laser desorption ionization-time of flight mass spectrometry[J].J Clin Microbiol,2013,51(7):2182-2194.
[15]Sparbier K,Weller U,Boogen C.Rapid detection of Salmonella sp.by means of a combination of selective enrichment broth and MALDI-TOFMS[J].Eur J Clin Microbiol Infect Dis,2012,31(5):767-773.
[2]Dieckmann R,Malorny B.Rapid screening of epidemiologically important Salmonella enterica subsp.enterica serovars by whole-cell matrix-assisted laser desorption ionization-time of flight mass spectrometry[J].Appl Environ Microbiol,2011,77(12):4136-4146.
[3]Huang CH,Huang L.Rapid species-and subspecies-specific level classification and identification of Lactobacillus casei group members using MALDI biotyper combined with clinprotools[J].J Dairy Sci,2018,101(2):979-991.
[4]Lawton SJ,Weis AM,Byrne BA,et al.Comparative analysis of campylobacter isolates from wild birds and chickens using MALDI-TOFMS,biochemical testing,and DNA sequencing[J].J Veter Diagnost Invest,2018,30(3):354-361.
[5]Pranada AB,Witt E,Bienia M,et al.Accurate differentiation of mycobacterium chimaera from mycobacterium intracellulare by MALDI-TOF MS analysis[J].J Med Microbiol,2017,66(5):670-677.
[6]Schlebusch S,Pric GR,Gallagher RL,et al.MALDI-TOF MS meets WGS in a VRE outbreak investigation[J].Eur J Clin Microbiol Infect Dis,2017,36(3):495-499.
[7]Ayeni FA,Andersen C,Lauritsen NN.Comparison of growth on mannitol salt agar,matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,VITEK?2 with partial sequencing of 16S rRNA gene for identification of coagulase-negative staphylococci[J].Microb Pathogen,2017,105:255-259.
[8]李修远,李亚楠,鲁辛辛.MALDI-TOF MS用于临床微生物检验中常见菌鉴定的应用探讨[J].临床检验杂志,2018,46(4):297-303.Li XY,Li YN,Lu XX.Discussion on the application of MALDI-TOF MSin the identification of common bacteria in clinical microbiological examination[J].J Clin Lab Sci,2018,46(4):297-303.
[9]佚名.MALDI-TOF MS病原体鉴定质量保证专家共识[J].中华检验医学杂志,2018,41(8):567-570.Anonymous.Expert consensus on MALDI-TOF MS pathogen identification quality assurance[J].Chin J Lab Med,2018,41(8):567-570.
[10]张雪英,叶红,李爱云,等.采用MALDI-TOF MS和全基因组测序鉴定和分析动弯杆菌属及其亲缘关系[J].中华微生物学和免疫学杂志,2018,38(6):446-450.Zhang XY,Ye H,Li AY,et al.Idention and analsis of genus acinetobacter and its relatives by MALDI-TOF MS and whole genome sequencing[J].Chin J Microbiol Immunol,2018,38(6):446-450.
[11]俞静,陈峰,李媛睿,等.应用MALDI-TOF MS技术快速鉴定布鲁菌[J].国际检验医学,2018,39(12):1443-1447.Yu J,Chen F,Li YR,et al.Rapidly identifition of Brucella by MALDI-TOF MS[J].Int Lab Med,2018,39(12):1443-1447.
[12]李颖,徐英春.MALDI-TOF MS在病原性丝状真菌鉴定中的应用进展[J].中国真菌学,2017,12(6):378-381.Li Y,Xu YC.Application progress of MALDI-TOF MS in identification of pathogenic filamentous fungi[J].Chin Mycol,2017,12(6):378-381.
[13]Li Y.Christiaens GCMLP,Gille JJP,et al.Non-invasive prenatal detection of achondroplasia in size-fractionated cell-free DNA by MALDI-TOF MSassay[J].Prenat Diagn,2007,27(1):11-17.
[14]Piseth Seng CA,Rolain JM,et al.Identification of rare pathogenic bacteria in a clinical microbiologylaboratory:impact of matrix-assisted laser desorption ionization-time of flight mass spectrometry[J].J Clin Microbiol,2013,51(7):2182-2194.
[15]Sparbier K,Weller U,Boogen C.Rapid detection of Salmonella sp.by means of a combination of selective enrichment broth and MALDI-TOFMS[J].Eur J Clin Microbiol Infect Dis,2012,31(5):767-773.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |