Effect of two kinds of bacterial pretreatment methods on the clinical isolate strains identification by MALDI-TOF MS

doi:10.3877/cma.j.issn.2095-5820.2018.01.008

Abstract

Abstract:

Objective

To evaluate the effect of different pretreatment methods on the identification of clinical bacterial isolates by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS).

Methods

Forty-three clinical isolates were collected and subcultured before treated by two different sample preparation methods including off-plate extraction method and direct on-plate method. And then the treated strains were identified by MALDI-TOF MS. The identification accuracy of the two methods and the peak of protein fingerprint were compared.

Results

Using the results of 16S rRNA or ITS RNA as reference standard, the identification rates of gram positive bacteria, gram negative bacteria and fungus treated by off-plate extraction method were 93.8%, 100% and 30.8%, respectively. And the identification rates of gram positive bacteria, gram negative bacteria and fungus treated by direct on-plate extraction method were 75.0%, 100% and 7.7%, respectively. The characteristic peak in the protein fingerprint obtained after extraction was more; the signal-to-noise ratio was higher and the reproducibility of protein fingerprint was better.

Conclusion

It is recommended that the off-plate extraction method for MALDI-TOF identification should be used, which is of great clinical significance for the establishment of a standard microbial detection process and the improvement of the accuracy of mass spectrometry identification.

Key words: Matrix-assisted laser desorption/ ionization time of flight mass spectrometry, Protein fingerprint, Pretreatment, Off-plate extraction method, Direct on-plate method

Weifan Cao, Zhao Song, Fengli Zhou, Xinchao Liu, Qingwei Ma, Fenglin Cao. Effect of two kinds of bacterial pretreatment methods on the clinical isolate strains identification by MALDI-TOF MS[J]. Chinese Journal of Clinical Laboratory Management(Electronic Edition), 2018, 06(01): 27-31.

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Figures/Tables 5

References 20

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菌株名称		株数	鉴定准确率（%）
菌株名称		株数	提取法	抹板法
革兰阳性细菌		?	?	?
?	粪肠球菌、表皮葡萄球菌、人葡萄球菌、小肠肠球菌、溶血葡萄球菌、屎肠球菌、铅黄肠球菌、解没食子酸链球菌、耐久肠球菌	12	100	100
?	鸟肠球菌	1	100	90
?	咽峡炎链球菌、金黄色葡萄球菌	2	100	80
?	头葡萄球菌	1	20	40
革兰阴性细菌		?	?	?
?	克氏柠檬酸杆菌、阴沟肠杆菌、铜绿假单胞菌、产酸克雷伯氏菌、产气肠杆菌、醋酸钙不动杆菌、醋酸钙不动杆菌－鲍曼不动杆菌复合体、弗劳地柠檬酸杆菌、大肠埃希氏菌、肺炎克雷伯氏菌、豚鼠/嗜水/简氏气单胞复合体、卡塔莫拉氏菌	14	100	100
真菌		?	?	?
?	近平滑念珠菌	1	100	90
?	粉状毕赤酵母	1	100	80
?	产朊假丝酵母	1	100	30
?	季也蒙毕赤酵母	1	100	10
?	季也蒙毕赤酵母	1	90	90
?	异常毕赤酵母	1	90	30
?	酿酒酵母	1	90	10
?	费比恩毕赤酵母	1	90	0
?	东方伊萨酵母	1	70	30
?	解脂耶氏酵母	1	70	0
?	白色念珠菌	1	60	100
?	白色念珠菌	1	60	80
?	东方伊萨酵母	1	40	30

菌株名称		株数	鉴定准确率（%）
菌株名称		株数	提取法	抹板法
革兰阳性细菌		?	?	?
?	粪肠球菌、表皮葡萄球菌、人葡萄球菌、小肠肠球菌、溶血葡萄球菌、屎肠球菌、铅黄肠球菌、解没食子酸链球菌、耐久肠球菌	12	100	100
?	鸟肠球菌	1	100	90
?	咽峡炎链球菌、金黄色葡萄球菌	2	100	80
?	头葡萄球菌	1	20	40
革兰阴性细菌		?	?	?
?	克氏柠檬酸杆菌、阴沟肠杆菌、铜绿假单胞菌、产酸克雷伯氏菌、产气肠杆菌、醋酸钙不动杆菌、醋酸钙不动杆菌－鲍曼不动杆菌复合体、弗劳地柠檬酸杆菌、大肠埃希氏菌、肺炎克雷伯氏菌、豚鼠/嗜水/简氏气单胞复合体、卡塔莫拉氏菌	14	100	100
真菌		?	?	?
?	近平滑念珠菌	1	100	90
?	粉状毕赤酵母	1	100	80
?	产朊假丝酵母	1	100	30
?	季也蒙毕赤酵母	1	100	10
?	季也蒙毕赤酵母	1	90	90
?	异常毕赤酵母	1	90	30
?	酿酒酵母	1	90	10
?	费比恩毕赤酵母	1	90	0
?	东方伊萨酵母	1	70	30
?	解脂耶氏酵母	1	70	0
?	白色念珠菌	1	60	100
?	白色念珠菌	1	60	80
?	东方伊萨酵母	1	40	30

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