基质辅助激光解析电离飞行时间质谱快速鉴定猪链球菌分离株  

Rapid identification of Streptococcus suis isolates by matrix assisted laser desorption ionization time of flight mass spectrometry

作　　者：谈忠鸣[1] 周伟忠[1] 董晨[1] 钱慧敏[1] 洪捷[1] 周璐[1] 孔筱筱 胡建利[1] 鲍倡俊[1] TAN Zhong-ming;ZHOU Wei-zhong;DONG Chen;QIAN Hui-ming;HONG Jie;ZHOU Lu;KONG Xiao-xiao;HU Jian-li;BAO Chang-jun(NHC Key Laboratory of Enteric Pathogenic Microbiology,Jiangsu Provincial Center for Disease Control and Prevention,Jiangsu Nanjing 210009,China)

机构地区：[1]国家卫生健康委员会肠道病原微生物重点实验室,江苏省疾病预防控制中心,江苏南京210009

出　　处：《江苏预防医学》2020年第6期603-605,共3页Jiangsu Journal of Preventive Medicine

基　　金：国家科技重大专项(2018ZX10714-002);江苏省医学重点学科(实验室)(ZDXKA2016008);江苏省疾病预防控制中心“科教强业”工程青年人才(JKRC2016005)。

摘　　要：目的评估基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)技术在猪链球菌快速鉴定中的可行性。方法采用MALDI-TOF-MS法,对江苏省分离的40株猪链球菌及感染后临床症状类似的肺炎链球菌、猩红热链球菌、牛链球菌、金黄色葡萄球菌、流感嗜血杆菌、肺炎克雷伯菌分离株进行分析,评估方法的特异性。分别在脱纤维羊血平板、普通营养琼脂平板及猪链球菌选择性平板上培养24、48、72、120h,鉴定后对结果进行描述分析。结果猪链球菌分离株40株主要为血清2型(占95.0%),分离自病人标本(占75.0%)。以MALDI-TOF-MS法分析,主要离子峰为2667.450、4133.787、4421.894、5968.847、8269.519、9335.019、10990.488、15055.203Da,峰谱稳定,鉴定准确率为100.0%。肺炎链球菌、猩红热链球菌、牛链球菌、金黄色葡萄球菌、流感嗜血杆菌、肺炎克雷伯菌分离株鉴定结果均为阴性。在3种平板上培养24、48、72h,猪链球菌分离株均能被MALDI-TOF-MS准确鉴定。结论建立的MALDI-TOF-MS法具有快速、精准、成本低的优点,可替代Vitek 2、PCR等方法或作为相互验证方法,在有条件的机构应用。Objective To evaluate the feasibility of matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS)for fast Streptococcus suis identification.Methods A total of 40 strains of Streptococcus suis and Streptococcus pneumoniae,Streptococcus scarlet,Streptococcus bovis,Staphylococcus aureus,Haemophilus influenzae and Klebsiella pneumoniae with similar clinical symptoms upon infection were analyzed by MALDI-TOF-MS.All isolates were cultured on the defibrinated sheep blood plate,ordinary nutrient agar plate and Streptococcus suis selective plate for 24,48,72 and 120 h,respectively.The results were described and analyzed after identification.Results The 40 strains of Streptococcus suis were mainly serotype 2(accounted for 95.0%)and isolated from patients(accounted for 75.0%).The main ion peaks were 2667.450,4133.787,4421.894,5968.847,8269.519,9335.019,10990.488,15055.203 Da.The spectrum was stable and consistant,the identification accuracy was 100.0%.All Streptococcus pneumoniae,Streptococcus scarlet,Streptococcus bovis,Staphylococcus aureus,Haemophilus influenzae and Klebsiella pneumoniae isolated strains showed negative results using the method for identificaiton.After cultured on 3 kinds of plates for 24,48 and 72 h,all Streptococcus suis isolates could be accurately identified by MALDI-TOF-MS.Conclusion The established MALDI-TOF-MS method is fast,accurate,with lowcost,which can replace Vitek 2 and PCR methods or serve as complement method and promoted in qualified facilities.

关 键 词：猪链球菌 基质辅助激光解析电离飞行时间质谱 快速鉴定 

分 类 号：R378.1[医药卫生—病原生物学]