[1]范 宁a,段雪红a,杨 瑜a,等.血培养阳性标本分离胶促凝管联合SDS 前处理与固体培养基短时培养菌膜应用MALDI-TOF MS 快速鉴定最佳时间探讨[J].现代检验医学杂志,2022,37(05):132-138.[doi:10.3969/j.issn.1671-7414.2022.05.026]
　FAN Ninga,DUAN Xue-honga,YANG Yua,et al.Discussion on the Best Time of Blood Culture Positive Samples for Rapid Identification by MALDI-TOF MS Using Separation Gel Accelerating Tube Combined with SDS Pretreatment and Short-term Culture of Biofilm on Solid Medium[J].Journal of Modern Laboratory Medicine,2022,37(05):132-138.[doi:10.3969/j.issn.1671-7414.2022.05.026]
点击复制

血培养阳性标本分离胶促凝管联合SDS 前处理与固体培养基短时培养菌膜应用MALDI-TOF MS 快速鉴定最佳时间探讨()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年05期
页码:
132-138
栏目:
论著
出版日期:
2022-09-15

文章信息/Info

Title:
Discussion on the Best Time of Blood Culture Positive Samples for Rapid Identification by MALDI-TOF MS Using Separation Gel Accelerating Tube Combined with SDS Pretreatment and Short-term Culture of Biofilm on Solid Medium
文章编号:
1671-7414（2022）05-132-07
作者:
范 宁a; 段雪红a; 杨 瑜a; 王媛媛a; 鱼丽萍b; 谢立民a; 王 苗a
咸阳市第一人民医院a. 检验科；b. 神经内科，陕西咸阳 712000
Author(s):
FAN Ninga;  DUAN Xue-honga;  YANG Yua;  WANG Yuan-yuana;  YU Li-pingb;  XIE Li-mina;  WANG Miaoa
a.Department of Clinical Laboratory；b.Department of Neurology, Xianyang First People’s Hospital, Shaanxi Xianyang 712000, China
关键词:
血流感染; 基质辅助激光解析电离飞行时间质谱; 分离胶促凝管; 十二烷基硫酸钠; 血培养
分类号:
Q503；R446.5
DOI:
10.3969/j.issn.1671-7414.2022.05.026
文献标志码:
A
摘要:
目的　探讨血培养阳性标本分离胶促凝管联合十二烷基硫酸钠（sodium dodecyl sulfate，SDS）前处理与固体培养基短时培养菌膜应用基质辅助激光解析电离飞行时间质谱（matrix-assisted laser desorption ionization time of flightmass spectrometry，MALDI-TOF MS）快速鉴定的最佳时间。方法　收集2020 年8 月～ 2021 年1 月来源于咸阳市第一人民医院微生物室的243 瓶血培养阳性（单数菌）样本，采用MALDI-TOF MS 技术对分离胶促凝管联合SDS 前处理标本直接鉴定，同时对固体培养基短时培养（6h 内）菌膜鉴定，以传统培养（24h）方法鉴定结果为金标准，分析MALDI-TOF MS 快速鉴定血培养阳性菌株的最佳方法和时间。结果　243 瓶血培养阳性标本共鉴定出病原菌44 种，分离率前五位的细菌为大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌、金黄色葡萄球菌和表皮葡萄球菌。SDS 前处理法直接鉴定总符合率为75.72%（184/243），其中革兰阴性菌和革兰阳性菌符合率分别为84.97%（130/153）和60.00%（54/90）；固体培养基生长2h，4h 和6h 的菌膜鉴定总符合率依次为44.44%（108/243），72.02%（175/243）和90.53%（220/243），其中革兰阴性菌依次为59.48%（91/153），83.01%（127/153）和92.81%（142/153），革兰阳性菌依次为18.89%（17/90）,53.33%（48/90）和86.67%（78/90）。金黄色葡萄球菌SDS 前处理法直接鉴定和2h，4h 生长菌膜鉴定符合率均高于凝固酶阴性葡萄球菌（χ2=4.795，12.083, 9.035，均P ＜ 0.01）。革兰阴性菌SDS 前处理法直接鉴定和2h，4h 生长菌膜鉴定符合率均高于革兰阳性菌（χ2=17.880，37.808，24.758, 均P ＜ 0.01)。结论　MALDI-TOF MS 可对血流感染（bloodstreaminfection，BSI）主要病原菌进行快速直接鉴定，根据革兰染色结果联合固体培养基短时培养菌膜鉴定，可进一步缩短细菌鉴定时间，为临床抗感染治疗提供早期依据。
Abstract:
Objective　To explore the best time for rapid identification of blood culture positive samples by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) using separation gel coagulation promoting tube combined with sodium dodecyl sulfate（SDS）pretreatment and short-term culture of bacterial membrane in solid medium. Methods　243 bottles of blood culture positive (singular bacteria) samples from Department of Clinical Laboratory, Xianyang First People’s Hospital from August 2020 to January 2021 were collected. The samples pretreated by gel promoting tube combined with SDS were directly identified by MALDI-TOF MS technology. At the same time, the bacterial membrane was identified by short-term culture in solid medium (within 6h). The identification results of traditional culture (24h) method were taken as the gold standard to analyze the best method and time for rapid identification of blood culture positive strains by MALDI-TOF MS. Results　A total of 44 kinds of pathogenic bacteria were identified in 243 blood culture positive samples. The top five bacteria were Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus and Staphylococcus epidermidis. The total coincidence rate of direct identification by SDS pretreatment method was 75.72% (184 / 243), of which the coincidence rates of Gram-negative bacteria and Gram-positive bacteria were 84.97% (130 / 153) and 60.00% (54 / 90), respectively. The total coincidence rates of bacterial membrane identification in solid medium grown for 2h, 4h and 6h were 44.44% (108 / 243), 72.02% (175 / 243) and 90.53% (220 / 243), respectively. Among them Gram negative bacteria were successively 59.48% (91 / 153), 83.01% (127 / 153) and 92.81% (142 / 153), respectively, and Gram positive bacteria were successively 18.89% (17 / 90), 53.33% (48 / 90) and 86.67% (78 / 90), respectively. The coincidence rates between SDS pretreatment identification and identification of biofilm after growing for 2h and 4h for Staphylococcus aureus were higher than those of coagulase negative Staphylococcus (χ2=4.795, 12.083, 9.035，all P ＜ 0.01）.For Gram-negative bacteria were higher than those of Gram-positive bacteria, the differences were statistically significant (χ2=17.880，37.808，24.758, all P<0.01). Conclusion　MALDI-TOF MS can quickly and directly identify the main pathogens of blood flow infection. According to the results of Gram staining combined with short-term culture of bacterial membrane in solid medium, the identification time can be further shortened, the identification accuracy can be improved, and the needs of early clinical etiological diagnosis and timely intervention of blood flow infection can be basically met.

参考文献/References:

[1] 丁毅伟, 吴思敏, 韩志海. 血流感染细菌快速鉴定及药敏试验的方法学评价[J]. 解放军医学杂志, 2021,46(5): 468-473.
DING Yiwei, WU Simin, HAN Zhihai. Methodological evaluation of rapid identification and drug susceptibility testing of bloodstream infection bacteria [J]. Medical Journal of Chinese People’s Liberation Army ,2021,46(5):468-473.
[2] 侯伟伟, 江涟, 李冬.MALDI-TOF MS 直接鉴定血培养阳性病原的临床应用[J].检验医学,2021,36(4):424-429.
HOU Weiwei, JIANG Lian, LI Dong. Clinical application of Matrix-assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry in the direct detection of blood culture positive specimens [J].Laboratory Medicine, 2021,36 (4): 424-429.
[3] 张菲菲, 王启, 李荷楠, 等. Sepsityper Kit 试剂盒法或血清分离胶法联合MALDI-TOF MS 快速鉴定血培养阳性病原菌[J]. 中华微生物学和免疫学杂志,2018, 38(2):111-115.
ZHANG Feifei, WANG Qi, LI Henan, et a1.Rapid identification of pathogenic organisms positive for blood culture by MALDI-TOF mass spectrometry in combination with Sepsityper Kit or serum-separating gel tubes [J]. Chinese Journal of Microbiology and Immunology, 2018,38 (2): 111-115.
[4] 宋启飞, 刘敏雪, 李梦娇, 等. 质谱快速鉴定血培养阳性标本的方法研究[J]. 中国现代医学杂志,2018,28(35):23-26.
SONG Qifei, LIU Minxue, LI Mengjiao, et al.Establishment of a method of rapid identification of positive blood culture samples by MALDI-TOF MS [J].China Journal of Modern Medicine, 2018,28 (35): 23-26.
[5] 周春妹, 沈佳瑾, 黄声雷, 等. 评估SDS 在MALDITOFMS直接鉴定阳性血培养样本中的应用价值[J].检验医学,2018,33(3):228-232.
ZHOU Chunmei, SHEN Jiajin, HUANG Shenglei,et al.SDS for improving direct identification of microorganisms in positive blood culturing by MALDITOF MS [J]. Laboratory Medicine, 2018,33 (3): 228-232.
[6] BROYER P, PERROT N, ROSTAING H, et al. An automated sample preparation instrument to accelerate positive blood cultures microbial identification by MALDI-TOF mass spectrometry (Vitek?MS) [J].Front Microbiol,2018,9:911.
[7] KAYIN M, MERT B, AYDEMIR S, et a1. Comparison of rapid BACpro? II, Sepsityper? kit and in-house preparation methods for direct identification of bacteria from blood cultures by MALDI-TOF MS with and without Sepsityper? module analysis [J]. Eur J Clin Microbiol Infect Dis, 2019,38(11):2133-2143.
[8] CHEN Xinfei , HOU Xin , XIAO Meng , et al. Matrixassisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analysis for the identification of pathogenic microorganisms: A review[J]. Microorganisms, 2021, 9(7):1536.
[9] 吴彩霞, 高玉芳, 郭梦, 等. 基质辅助激光解析电离飞行时间质谱在血流感染快速鉴定中的应用[J]. 甘肃科学学报, 2019, 31(4)：66-69.
WU Caixia, GAO Yu f a n g , GUO Meng, e t al.Application of Matrix-Assisted Laser Desorption Lionization Time of Flight Mass Spectrometry in rapid identification of blood flow infection [J]. Journal of Gansu Sciences,2019,31(4):66-69.
[10] 陈峰, 李媛睿, 皇甫昱婵, 等. 分离胶促凝管联合MALDI-TOF MS 直接检测血培养阳性细菌[J]. 检验医学,2015,30(2):113-121.
CHEN Feng, LI Yuanrui, HUANGFU Yuchan, et al.Direct identification of microorganisms from blood culture by MALDI-TOF MS combined with separation gel tube [J]. Laboratory Medicine, 2015,30 (2): 113-121.
[11] 马坚, 俞万钧, 胡必杰, 等. 通过基质辅助激光解析电离飞行时间质谱系统直接快速鉴定阳性血培养[J]. 中华医院感染学杂志,2017,27(12):2676-2679.
MA Jian, YU Wanjun, HU Bijie, et al. Rapid method for direct identification of bacteria in blood culture broth using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry [J]. Chinese Journal of Nosocomiology,2017,27 (12): 2676-2679.
[12] 王军杰, 马冰, 李轶, 等. 质谱技术直接鉴定报阳血培养细菌可行性分析[J]. 检验医学,2020,35(7):716-720.
WANG Junjie, MA Bing, LI Yi, et al. Identification of bacteria directly by MALDI-TOF MS [J]. Laboratory Medicine, 2020,35 (7): 716-720.
[13] 方毅, 张景皓, 赵虎. 分离胶与快速培养富集法在MALDI-TOF MS 快速检测报阳血培养样本的方法学比较[J]. 老年医学与保健,2018,24(6):705-708, 761.
FANG Yi , Z H A N G J i n g h a o , ZHAO Hu.
Methodological comparison of separation gel tube and rapid culture in direct identification of bacteria from positive blood cultures by MALDI-TOF MS [J].Geriatrics & Health Care,2018,24 (6): 705-708, 761.
[14] 余佳佳, 李媛睿, 刘瑛. SDS 和SAP 前处理联合MALDI-TOF MS 在报阳血培养样本快速病原菌鉴定中的价值[J]. 检验医学,2021,36(8):864-868.
YU Jiajia, LI Yuanrui, LIU Ying. MALDI-TOF MS combined with SDS and SAP pretreatment for rapid identification of positive blood culture [J]. Laboratory Medicine,2021,36(8):864-868.
[15] MACHEN A, DRAKE T, WANG Y F. Same day identification and full panel antimicrobial susceptibility testing of bacteria from positive blood culture bottles made possible by a combined lysis-filtration method with MALDI-TOF VITEK mass spectrometry and the VITEK2 system [J]. PLoS One,2014 ,9(2):e87870.
[16] 谷钰峰, 李昱, 张晓录, 等. 基于MALDI-TOF MS与FCM 联合应用对临床血流感染病原体鉴定和药敏试验新方法的建立和应用[J]. 现代检验医学杂志,2018,33(4):20-24.
GU Yufeng, LI Yu, ZHANG Xiaolu, et al. A new method aimed to quickly identify pathogen and drug susceptibility test based on MALDI-TOF MS combined with FCM [J]. Journal of Modern Laboratory Medicine,2018,33(4):20-24.
[17] HAMILTON F, EVANS R, MACGOWAN A. The value of MALDI-TOF failure to provide an identification of Staphylococcal species direct from blood cultures and rule out Staphylococcus aureus bacteraemia : a post-hoc analysis of the RAPIDO trial [J]. Access Microbiol. 2020,3(2):000192.
[18] 林晓晖, 麦惠香, 简梦华, 等. 梅里埃VTIEKMS 质谱仪在甲氧西林耐药及敏感的金黄色葡萄球菌快速鉴定的应用研究[J]. 现代检验医学杂志,2020,35(5):121-123.
LIN Xiaohui, MAI Huixiang, JIAN Menghua, et al.Study on VTIEK MS mass spectrometer rapid identification of methicillin resistance and sensitive Staphylococcus aureus [J]. Journal of Modern Laboratory Medicine,2020,35(5):121-123.
[19] IDELEVICH EA, REISCHL U, BECKER K.
New microbiological techniques in the diagnosis of bloodstream infections [J]. Dtsch Arztebl Int,2018,115(49):822-832.
[20] 吴富炜, 王圆圆, 杨靖娴. 短时固相培养法联合MALDI-TOF MS 快速鉴定含活性炭阳性血培养标本[J]. 标记免疫分析与临床,2020,27(12):2176-2180.
WU Fuwei, WANG Yuanyuan, YANG Jingxian .The rapid identification of microorganisms from positive blood culture containing charcoal by MALDI-TOF MS after short-term incubation on solid medium[J]. Labeled Immunoassays and Clinical Medicine,2020,27(12):2176-2180.
[21] HA J, HONG S K, HAN G H, et al. Same-day identification and antimicrobial susceptibility testing of bacteria in positive blood culture broths using shortterm incubation on solid medium with the MicroFlex LT, Vitek-MS, and Vitek2 Systems [J]. Ann Lab Med.
2018,38(3):235-241.
[22] 周道红, 黎敏, 鲁卫平. 利用MALDI-TOF MS 快速鉴定经固体培养基短时培养的阳性血培养物中的病原菌[J]. 国际检验医学杂志,2018,39(22):2746-2749,2755.
ZHOU Daohong, LI Min, LU Weiping .Rapid identification of microorganisms from positive blood cultures by MALDI-TOF Mass Spectrometry following short-term incubation on solid medium [J]. International Journal of Laboratory Medicine, 2018,39(22):2746-2749,2755.
[23] 中国医疗保健国际交流促进会临床微生物与感染分会, 中华医学会检验医学分会临床微生物学组,中华医学会微生物学和免疫学分会临床微生物学组. 血液培养技术用于血流感染诊断临床实践专家共识[J]. 中华检验医学杂志,2022,45(2):105-121.
Society of Clinical Microbiology and Infection of China International Exchange and Promotion Association for Medical and Healthcare, Clinical Microbiology Group of the Laboratory Medicine Society of the Chinese Medical Association, Clinical Microbiology Group of the Microbiology and Immunology Society of the Chinese Medical Association .Chinese expert consensus on the clinical practice of blood culture in the diagnosis of bloodstream infection [J]. Chinese Journal of Laboratory Medicine,2022,45(2):105-121.
[24] YUAN Youhua, WANG Junjie, ZHANG Jiangfeng,et a1. Evaluation of an optimized method to directly identify bacteria from positive blood cultures using MALDI-TOF mass spectrometry [J]. J Clin Lab Anal,2020,34(4):e23119.

相似文献/References:

[1]翟华丽,刘艳丽,丁丽丽,等.2012～2014年医院血培养主要病原菌分布及耐药性分析[J].现代检验医学杂志,2015,30(03):104.[doi:10.3969/j.issn.1671-7414.2015.03.030]
　ZHAI Hua-li,LIU Yan-li,DING Li-li,et al.Distribution and Drug Resistance of Main Pathogens Isolated from Blood Culture from 2012 to 2014[J].Journal of Modern Laboratory Medicine,2015,30(05):104.[doi:10.3969/j.issn.1671-7414.2015.03.030]
[2]谷钰峰,李 昱,张晓录,等.基于MALDI-TOFMS与FCM联合应用对临床血流感染病原体鉴定和药敏试验新方法的建立和应用[J].现代检验医学杂志,2018,33(04):2.[doi:10.3969/j.issn.1671-7414.2018.04.006]
　GU Yu-feng,LI Yu,ZHANG Xiao-lu,et al.A New Method Aimed to Quickly Identify Pathogen and Drug Susceptibility Test Based on MALDI-TOF MS Combined with FCM[J].Journal of Modern Laboratory Medicine,2018,33(05):2.[doi:10.3969/j.issn.1671-7414.2018.04.006]
[3]张 戈,张小江,胡继红,等.国产全自动血培养系统及配套血培养瓶性能评估[J].现代检验医学杂志,2018,33(06):132.[doi:10.3969/j.issn.1671-7414.2018.06.034]
　ZHANG Ge,ZHANG Xiao-jiang,HU Ji-hong,et al.Performance Evaluation of Domestic LABSTAR120 Automated Blood Culture System and Its Blood Culture Bottles[J].Journal of Modern Laboratory Medicine,2018,33(05):132.[doi:10.3969/j.issn.1671-7414.2018.06.034]
[4]王 晶a,马 娟a,范 云a,等.2015～2017年陕西省人民医院临床血流感染病原菌的分布及耐药性分析[J].现代检验医学杂志,2019,34(04):87.[doi:10.3969/j.issn.1671-7414.2019.04.021]
　WANG Jinga,MA Juana,FAN Yuna,et al.Distribution and Drug Resistance of Pathogenic Bacteria of Blood Culture in Shaanxi Provincial People's Hospital form 2015 to 2017[J].Journal of Modern Laboratory Medicine,2019,34(05):87.[doi:10.3969/j.issn.1671-7414.2019.04.021]
[5]马梦亭,王凤超.临床患者血流真菌感染的实验诊断及治疗研究进展[J].现代检验医学杂志,2019,34(04):161.[doi:10.3969/j.issn.1671-7414.2019.04.040]
　MA Meng-ting,WANG Feng-chao.Advances in Experimental Diagnosis and Treatment of Blood Flow Fungal Infection in Clinical Patients MA Meng-ting,WANG Feng-chao[J].Journal of Modern Laboratory Medicine,2019,34(05):161.[doi:10.3969/j.issn.1671-7414.2019.04.040]
[6]关佳灏,赵　海,王　翠,等.2018年陕西省人民医院全自动血培养仪及配套鉴定系统分离病原菌的实验室特征及耐药分析[J].现代检验医学杂志,2019,34(06):113.[doi:10.3969 ／ j.issn.1671-7414.2019.06.028]
　GUAN Jia-hao,ZHAO Hai,WANG Cui,et al.Laboratory Characteristics and Drug Resistance Analysis of Pathogenic Bacteria Isolated by Automatic Blood Culture Instrument and Matching Idetification System in Shaanxi Provincial People’s Hospital in 2018[J].Journal of Modern Laboratory Medicine,2019,34(05):113.[doi:10.3969 ／ j.issn.1671-7414.2019.06.028]
[7]任继欣,郭彦言.外周血HBP，IL-6，CD64 指数，PCT，CRP 和SAA 水平检测在血流感染诊断中的应用价值研究[J].现代检验医学杂志,2020,35(02):56.[doi:10.3969/j.issn.1671-7414.2020.02.017]
　REN Ji-xin,GUO Yan-yan.Application Value of Peripheral Blood HBP, Interleukin-6, CD64 Index, PCT,CRP ，SAA Level Detection in Blood Stream Infection Diagnosis[J].Journal of Modern Laboratory Medicine,2020,35(05):56.[doi:10.3969/j.issn.1671-7414.2020.02.017]
[8]屈晨虹a,丁　雨b,史伟峰b.MADLI-TOF MS 和PhoenixTM-100 两种分析仪鉴定肠杆菌科细菌一致性评价[J].现代检验医学杂志,2020,35(02):97.[doi:10.3969 ／ j.issn.1671-7414.2020.02.027]
　QU Chen-honga,DING Yub,SHI Wei-fengb.Consistency of MADLI-TOF MS and PhoenixTM-100 to IdentifyEnterobacteriaceae Bacteria[J].Journal of Modern Laboratory Medicine,2020,35(05):97.[doi:10.3969 ／ j.issn.1671-7414.2020.02.027]
[9]何家花,余成强,李步荣.血流感染病原学分析及与相关炎性因子的应用价值研究[J].现代检验医学杂志,2020,35(04):139.[doi:10.3969/j.issn.1671-7414.2020.04.035]
　HE Jia-hua,YU Cheng-qiang,LI Bu-rong.Pathogen Analysis of Bloodstream Infection and Study on the Application Value of Related Inflammatory Factors[J].Journal of Modern Laboratory Medicine,2020,35(05):139.[doi:10.3969/j.issn.1671-7414.2020.04.035]
[10]吕梅,杨旭,和平安,等.老年患者由产气荚膜梭菌引起肺部和血流感染的临床和实验室分析及相关文献复习[J].现代检验医学杂志,2020,35(05):154.[doi:10.3969/j.issn.1671-7414.2020.05.040]
　L? Mei,YANG Xu,HE Ping-an,et al.Clinical and Laboratory Analysis and Literature Review of Pulmonary and Bloodstream Infections Caused by Clostridium Perfringens in Elderly Patients[J].Journal of Modern Laboratory Medicine,2020,35(05):154.[doi:10.3969/j.issn.1671-7414.2020.05.040]

备注/Memo

备注/Memo:
基金项目：咸阳市2020 年重点研发计划（2020k02-106）。
作者简介：范宁（1973-），女，本科，主任技师，主要从事临床微生物检验及细菌耐药性研究，E-mail：330464724@qq.com。
通讯作者：杨瑜（1984-），女，主管技师，主要从事临床微生物及免疫学检验工作，E-mail：2191487760@qq.com。
更新日期/Last Update: 2022-09-15