一种β-内酰胺类抗生素的酶热检测方法

• The penicillinase thermistor biosensor（Penicillinase sensor） was developed for the rapid monitoring of blood penem antibiotics concentration and rapid identification of extraneous penicillinase in milk on site.However, the wide application of the penicillinase thermistor biosensor was limited due to its intrinsic poor activity to hydrolyze cephem and carbapenem antibiotics.The recently identified carbapenemase New Delhi metallo-beta-lactamase 1（NDM-1） is able to hydrolyze all commercially available β-lactam antibiotics in high efficacy.We coupled the NDM-1 and the enzymatic thermistor biosensor to develop a NDM-1 thermistor biosensor（NDM-1 sensor） by the installment of the enzymatic thermistor with an enzyme column loaded with NDM-1 conjugated CPG beads.The NDM-1 sensor shows high response activity to Piperacillin（PIP）,Ceftriaxone（CTRX）, and Meropenem（MEM）, and the response activity of the NDM-1 sensor to these three β-lactam antibiotics are all Zn2+ dependent.Moreover, the response activity of the NDM-1 sensor to Penicillin G（P）, PIP, Cefazolin（CZO）, CTRX, Cefepime（FEP） and MEM all linearly correlated with antibiotic concentration from 31.25 to 1 000 mg/L.Within pH from 6.0 to 8.0, the optimal response activity of the NDM-1 sensor to P,PIP, CZO, CTRX and FEP are found at pH 6.5, while the optimal response activity of the NDM-1 sensor to MEM is found at pH8.0.These data indicate that the featured activity of NDM-1 was well maintained after conjugation on CPG beads, and NDM-1 sensor is capable to quantitate three classes of β-lactam antibiotics including penem, cephem and carbapenem within a wide concentration range.

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

β-lactam antibiotics

enzyme thermistor

NDM-1

blood antibiotic concentration

antibiotics resistance

Publikations- och innehållstyp

art (ämneskategori)

ref (ämneskategori)