检验医学
檢驗醫學
검험의학
LABORATORY MEDICINE
2015年
2期
113-121
,共9页
陈峰%李媛睿%皇甫昱婵%陶晓勤%刘瑛
陳峰%李媛睿%皇甫昱嬋%陶曉勤%劉瑛
진봉%리원예%황보욱선%도효근%류영
分离胶促凝管%基质辅助激光解析电离飞行时间质谱%血培养
分離膠促凝管%基質輔助激光解析電離飛行時間質譜%血培養
분리효촉응관%기질보조격광해석전리비행시간질보%혈배양
Separation gel tube%Matrix-assisted laser desorption/ionization-time of flight mass spectrometry%Blood culture
目的:建立用分离胶促凝管联合基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS )直接检测血培养阳性待测菌的方法并评估其鉴定符合率。方法共收集阳性血培养491例,利用分离胶促凝管直接从血培养瓶中富集并提纯菌体,采用 MALDI-TOF MS 对待测菌进行菌种鉴定,同时对报阳性血培养瓶进行转种培养,纯菌落用 Vitek 2 Compact 全自动微生物分析系统(简称 Vitek 2 Compact)进行鉴定。若两者鉴定结果不一致,则以基因测序结果予以确证。结果491例阳性血培养瓶鉴定出的待测菌包括30个属和64个种。在462例单菌株感染血培养瓶中,菌株的种、属鉴定符合率分别为73.6%和3.7%。其中175例革兰阴性菌的种、属鉴定率分别为84.0%(147例)和2.9%(5例);251例革兰阳性菌的种、属鉴定率分别为75.3%(189例)和4.4%(11例);36例念珠菌的种、属鉴定率分别为19.4%(7例)和2.8%(1例)。在29例复数菌感染血瓶中,鉴定出其中一种细菌的种、属鉴定率分别为79.3%(23例)和10.3%(3例)。血流感染中常见病原菌的菌种鉴定符合率达83.3%~96.9%。结论本研究建立了分离胶促凝管联合 MALDI-TOF MS 直接检测血培养阳性待测菌的方法,与传统的培养鉴定方法相比,对血流感染中主要病原菌的鉴定符合率较高,且方法快速、简便,成本低廉,适合在临床微生物实验室中推广应用。
目的:建立用分離膠促凝管聯閤基質輔助激光解析電離飛行時間質譜(MALDI-TOF MS )直接檢測血培養暘性待測菌的方法併評估其鑒定符閤率。方法共收集暘性血培養491例,利用分離膠促凝管直接從血培養瓶中富集併提純菌體,採用 MALDI-TOF MS 對待測菌進行菌種鑒定,同時對報暘性血培養瓶進行轉種培養,純菌落用 Vitek 2 Compact 全自動微生物分析繫統(簡稱 Vitek 2 Compact)進行鑒定。若兩者鑒定結果不一緻,則以基因測序結果予以確證。結果491例暘性血培養瓶鑒定齣的待測菌包括30箇屬和64箇種。在462例單菌株感染血培養瓶中,菌株的種、屬鑒定符閤率分彆為73.6%和3.7%。其中175例革蘭陰性菌的種、屬鑒定率分彆為84.0%(147例)和2.9%(5例);251例革蘭暘性菌的種、屬鑒定率分彆為75.3%(189例)和4.4%(11例);36例唸珠菌的種、屬鑒定率分彆為19.4%(7例)和2.8%(1例)。在29例複數菌感染血瓶中,鑒定齣其中一種細菌的種、屬鑒定率分彆為79.3%(23例)和10.3%(3例)。血流感染中常見病原菌的菌種鑒定符閤率達83.3%~96.9%。結論本研究建立瞭分離膠促凝管聯閤 MALDI-TOF MS 直接檢測血培養暘性待測菌的方法,與傳統的培養鑒定方法相比,對血流感染中主要病原菌的鑒定符閤率較高,且方法快速、簡便,成本低廉,適閤在臨床微生物實驗室中推廣應用。
목적:건립용분리효촉응관연합기질보조격광해석전리비행시간질보(MALDI-TOF MS )직접검측혈배양양성대측균적방법병평고기감정부합솔。방법공수집양성혈배양491례,이용분리효촉응관직접종혈배양병중부집병제순균체,채용 MALDI-TOF MS 대대측균진행균충감정,동시대보양성혈배양병진행전충배양,순균락용 Vitek 2 Compact 전자동미생물분석계통(간칭 Vitek 2 Compact)진행감정。약량자감정결과불일치,칙이기인측서결과여이학증。결과491례양성혈배양병감정출적대측균포괄30개속화64개충。재462례단균주감염혈배양병중,균주적충、속감정부합솔분별위73.6%화3.7%。기중175례혁란음성균적충、속감정솔분별위84.0%(147례)화2.9%(5례);251례혁란양성균적충、속감정솔분별위75.3%(189례)화4.4%(11례);36례념주균적충、속감정솔분별위19.4%(7례)화2.8%(1례)。재29례복수균감염혈병중,감정출기중일충세균적충、속감정솔분별위79.3%(23례)화10.3%(3례)。혈류감염중상견병원균적균충감정부합솔체83.3%~96.9%。결론본연구건립료분리효촉응관연합 MALDI-TOF MS 직접검측혈배양양성대측균적방법,여전통적배양감정방법상비,대혈류감염중주요병원균적감정부합솔교고,차방법쾌속、간편,성본저렴,괄합재림상미생물실험실중추엄응용。
Objective To evaluate the direct identification of microorganisms from blood culture by a separation gel tube-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS ) and its identification rate.Methods A total of 491 cases with positive blood culture were collected.The bacteria were enriched and purified directly from blood culture bottle by separation gel tube,and then the bacteria were identified by MALDI-TOF MS.Traditional culture and identification processes were performed simultaneously,and the cultured pure colonies were identified by Vitek 2 Compact automated microbial analysis system (Vitek 2 Compact).The identification of bacterial isolates by MALDI-TOF MS was compared with that of traditional biochemical testing,and discrepancies were resolved by gene sequencing.Results A total of 491 positive cultures were determined,representing 30 genera and 64 species or groups.The method demonstrated 73.6% and 3.7% concordance to species and genus identification rates in monomicrobial culture (462 cases).The correct species and genus identification rates were 84.0%(1 47 cases) and 2.9%(5 cases)in Gram-negative bacteria(1 75 cases),were 75.3%(1 89 cases)and 4.4%(1 1 cases)in Gram-positive bacteria(251 cases),and were 1 9.4%(7 cases)and 2.8% (1 case)in Candida species(36 cases).In 29 cases of complex bacterial infection,the correct species and genus identifications rates in one of the multiple bacteria were 79.3% (23 cases)and 1 0.3% (3 cases).Species identification rates of bloodstream infection with common pathogens were 83.3%-96.9%.Conclusions This separation gel tube-based MALDI-TOF MS for the direct identification of blood culture pathogens is established.The identification rate of common pathogens in bloodstream infection by MALDI-TOF MS is higher compared with that by traditional culture identification processes.In addition,it is rapid,easy and cost-effective,and it is a recommendable method for clinical microbiology laboratories.