A minisonicator to rapidly disrupt bacterial spores for DNA analysis.
نویسندگان
چکیده
Concerns about the use of anthrax spores as a weapon of mass destruction have motivated the development of portable instruments capable of detecting and monitoring a suspected release of the agent. Optimal detection of bacterial spores by PCR requires that the spores be disrupted to make the endogenous DNA available for amplification. The entire process of spore lysis, PCR, and detection can take several hours using conventional methods and instruments. In this report, a minisonicator and prototype spore lysis cartridge were built to disrupt Bacillus spores in 30 s for rapid, real-time PCR analysis. Utilization of the minisonicator improved PCR analysis by decreasing the limit of detection, reducing the time of detection, and increasing the signal amplitude. Total time of spore disruption and detection using the minisonicator and a microchip PCR instrument was less than 15 min.
منابع مشابه
Microfluidic sonicator for real-time disruption of eukaryotic cells and bacterial spores for DNA analysis.
Biologic agent screening is a three-step process: lysis of host cell membranes or walls to release their DNA, polymerase chain reaction to amplify the genetic material and screening for distinguishing genetic signatures. Macrofluidic devices commonly use sonication as a lysis method. Here, we present a piezoelectric microfluidic minisonicator and test its performance. Eukaryotic human leukemia ...
متن کاملSmall-Scale DNA Sample Preparation Method for Field PCR Detection of Microbial Cells and Spores in Soil.
Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial en...
متن کاملDetection of Paenibacillus larvae larvae spores in honey and diseased larvae Samples by culture and PCR
American foulbrood (AFB) is the most serious brood disease of the honey bee. Traditional methods are reliable but rather slow simply because they are based on biochemical, morphological and physiological identification of cultivated isolates. The aim of this study is the detection of Paenibacillus larvae larvae spores in honey and diseased larvae samples by culture and PCR. Therefore 54 samp...
متن کاملجذب اسپور های باکتریایی از هوا توسط خاکستر استخوان
Background and aims : Microorganisms are one of the most important sources of contamination in indoor air and occupational environments especially in hospitals. The aim of this study was to evaluate the adsorption characteristics of bone char to adsorb B. Subtilis in the air. Methods: In this study B.subtilis spores were used as a model of bacterial spores. Also bone char was used as an ads...
متن کاملAmplicon sequencing for the quantification of spoilage microbiota in complex foods including bacterial spores
BACKGROUND Spoilage of food products is frequently caused by bacterial spores and lactic acid bacteria. Identification of these organisms by classic cultivation methods is limited by their ability to form colonies on nutrient agar plates. In this study, we adapted and optimized 16S rRNA amplicon sequencing for quantification of bacterial spores in a canned food matrix and for monitoring the out...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Analytical chemistry
دوره 71 19 شماره
صفحات -
تاریخ انتشار 1999