METHOD OF DIRECTLY DETECTING ACTIVE FORM OF BETA-LACTAM ANTIBIOTICS HYDROLASE USING MASS SPECTROMETRY

• Main Authors: KO, Ji Hye, ROH, Jae Woo, YOON, Eun-Jeong, LEE, Eun Hee, HWANG, Dong Hwi, JI, In Jung, PARK, Kye Shin, PARK, Joon Sang, AHN, Jin Sung
• Format: Patent
• Language: English; French; Korean
• Published: 06.02.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; TESTING; VINEGAR; WINE

The present invention relates to a method of directly detecting, via mass spectrometry, whether or not bacteria in a sample have resistance to an antibiotic agent, and a detection kit for use in the method. More specifically, the present invention relates to a method and kit, which enables a direct determination of whether or not the bacteria are resistant to the antibiotic agent through direct detection of an antibiotic agent hydrolase secreted by the bacteria resistant to the antibiotic agent. According to the present invention, the resistance of a bacterial strain to an antibiotic agent can be confirmed immediately on the spot in a very convenient manner. In particular, the present invention provides: a method that, without undergoing complicated pretreatment processes such as protein digestion, or complicated identification processes involving calibrating and combining obtained results, can confirm resistance to the antibiotic agent within ten minutes or so, the process which typically takes a few days; and a simple detection kit for use in the method. La présente invention concerne un procédé permettant de détecter directement, par spectrométrie de masse, si des bactéries dans un échantillon présentent une résistance à un agent antibiotique, ainsi qu'un kit de détection destiné à être utilisé dans le procédé. Plus précisément, la présente invention concerne un procédé et un kit qui permettent de déterminer directement si les bactéries sont résistantes à l'agent antibiotique par la détection directe d'une hydrolase d'agent antibiotique secrétée par les bactéries résistantes à l'agent antibiotique. Selon la présente invention, la résistance d'une souche bactérienne à un agent antibiotique peut être confirmée immédiatement sur place d'une manière très commode. En particulier, la présente invention concerne : un procédé qui, sans subir un processus de prétraitement compliqué tel que la digestion de protéines ou des processus d'identification compliqués impliquant l'étalonnage et la combinaison de résultats obtenus, peut confirmer la résistance à l'agent antibiotique en dix minutes environ, pour un processus prenant habituellement quelques jours ; et un kit de détection simple destiné à être utilisé dans le procédé. 본 발명은 시료 중 포함된 미생물이 항생제에 대하여 내성을 갖는 미생물인지 여부를 질량분석법을 통하여 직접적으로 검출해내는 방법 및 이에 사용되는 검출용 키트에 관한 것이다. 보다 구체적으로는, 항생제에 내성인 미생물이 분비하는 항생제 가수분해 효소를 직접적으로 검출함으로써, 해당 미생물이 항생제에 대하여 내성인지 여부를 직접적으로 판별할 수 있는 방법 및 키트에 관한 것이다. 본 발명에 의하면 특정 균주의 항생제 내성 여부를 매우 간편하게 현장에서 즉시 확인할 수 있다. 특히, 단백질 분해 등의 복잡한 전처리 과정을 거치지 않으며, 얻어진 결과를 캘리브레이션하여 조합하는 복잡한 확인과정을 거치지 않을 수 있어, 기존 수일이 걸리던 항생제 내성 여부를 단 십여분 만에 간편하게 확인할 수 있는 방법, 및 이에 이용되는 간편 진단용 키트를 얻을 수 있다.