Detection of Staphylococcus aureus by a lambda exonuclease based piezoelectric quartz crystal microbalance DNA sensor
- VernacularTitle:核酸外切酶-压电石英DNA传感器检测金黄色葡萄球菌的实验研究
- Author:
Junfu HUANG
;
Han XIA
;
Qing HUANG
;
Weiling FU
- Publication Type:Journal Article
- Keywords:
quartz crystal microbalance;
biosensor;
lambda exonuclease;
Staphylococcus aureus
- From:Journal of Third Military Medical University
2003;0(13):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a new type of piezoelectric quartz crystal microbalance DNA sensor based on lambda exonuclease to detect Staphylococcus aureus (S. aureus) and optimize the main detection conditions. Methods After the DNA was extracted from the stain of S. aureus and the target fragment was amplified with self-designed universal primers, the PCR products were treated with Lambda exonuclease and visualized by agarose gel electrophoresis with ethidium bromide followed by a gel documentation system analysis. Then the products with or without exonuclease digestion were added into our piezoelectric quartz crystal microbalance DNA sensor for hybridization. The temperature and time of hybridization were optimized respectively. The specificity and sensitivity of our detection system were evaluated. Results The optimal temperature for hybridization was 35℃ and the optimal time was 60 min. The sensitivity of the lambda exonuclease based DNA sensor was better than that of common DNA sensor. The lowest detection limit of this new type quartz crystal microbalance system was 1.0?104 CFU/ml. Conclusion The lambda exonuclease based quartz crystal microbalance system we designed has the advantages of high efficiency in hybridization, easy to operate, and good response performance. Thus it can be applied to detect S. aureus infections.
