Flight time mass spectrometer identification microbial introduction
Flight time mass spectrometer identification microbial introduction Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF) fingerprinting is a fast and reliable method for microbial classification and identification, which can be widely used in clinical diagnosis, environmental and classification research or food processing quality control. BioTyperTM MALDI-TOF mass spectrometry fingerprints allow researchers to identify unknown bacteria, yeasts and molds in minutes. Repeatability This method has proven to be very stable over a wide range of conditions. The different composition of the growth medium had a very small effect on the peak pattern distribution; in the range from 4000 to 12000 Da, almost no effect of the medium was observed. The medium attached to the clone had no effect on the peak pattern. Similarly, the growth state of the cells has no effect on the peak pattern. Cells in the slow growth phase have similar patterns to cells in the logarithmic growth phase, plateau or death phase. Software analysis The analytical software provided by the manufacturer integrates all the functions of the identified and classified processed mass spectrograms. These processing parameters are user-definable and the result is a specialized list of peaks. To identify unknown microorganisms, pattern matching is accomplished by comparing the resulting peak list to a library of spectra containing a large number of species characteristics. The signature library is generated by several measurements of known bacterial species and strains under subtle conditions. The characteristic peak information is then extracted, and the good average is obtained by measuring 20 spectra. The software automatically generates a peak list from the entire spectral set and extracts typical peaks that appear in a certain number of spectra from a genus source. The use of BioTyper provides an excellent method for the identification of traditional microbiology laboratories in many fields, such as environmental research, food and water control and medical diagnostics. Its rapid and stable sample preparation and measurement makes it ideal for routine use. And high-throughput applications. Gynaecology Products,Cervical Scraper,Disposable Endometrial Cannula,Gynaecological Brush Luck Medical Consumables Co.,LIMITED , https://www.luckmedical.com
The general workflow for microbial expression profiling is a linear path. Starting with a monoclonal or other biological material, samples can be analyzed in minutes. Automatic spectral acquisition of each sample can be completed in a matter of seconds, and seamless transfer of data to specialized authentication software can be achieved. This technology is therefore an excellent alternative to classical microbial identification and classification techniques, requiring minimal sample processing and cost.
MALDI-TOF Determination Most simple sample analysis begins with placing a small amount of biological material directly onto the MALDI-TOF target. The starting material can be a component of a single or liquid medium centrifuge. The microbial film is covered with a matrix (α-cyano-4-hydroxycinnamic acid; HCCA). Mass spectrograms were acquired in a linear positive mode using a MALDI-TOF mass spectrometer with maximum frequency (20-200 Hz, depending on the instrument). The measurement quality of the spectrum is 2000-20000Da. Automatic spectral acquisition can be performed using Bluker's 'autoExecute' software with fuzzy control of laser intensity.
In addition, the mass spectrograms acquired on different MALDI-TOF instruments are highly comparable after sample preparation and measurement under standard conditions. The spectra measured on the same sample target were virtually identical across 3 different instruments. Therefore, spectra from different MALDI-TOF mass spectrometers can be used to build a truly reliable database.
The surprising reproducibility of this methodology is based on highly abundant protein measurements of stable expression, such as ribosomal proteins. Spectrograms can also be observed in the mass range of 2000 to 20000 Da with few metabolites. Bacterial spores can produce distinct peak patterns compared to live cells, and these "spore spectra" are also reproducible.
Unknown microorganisms are identified by comparison of their respective peak lists with databases. A match value is generated based on the determined mass and the correlation of their intensities, which are used to sort the results. To improve the accuracy of database searches, BioTyper can correct peak mass shifts with an advanced recalibration algorithm. After peak extraction, an initial acceptable error window can be set and a desired adjustment result can be obtained. The software can perform a new peak list calibration of the known peak list within an adjustable range. Therefore, even spectra with a mass deviation of 5000 ppm can still be successfully identified. This feature makes BioTyper identification exceptionally reliable and accurate.
To reduce duplication, BioTyper provides many features such as clustering and generating family trees. Based on the similarity score, a tree diagram can be created. Moreover, an unregulated multivariate analysis based on factor analysis is possible. Using calculated feature components, multiple clustering algorithms and visualization methods are all achievable.
This method is also very useful for phylogenetic analysis. MALDI-TOF mass spectrometry expression profiling produces the same family tree as classical 16S ribosomal DNA sequencing. Because ribosomal proteins are highly abundant and very stable, observation of protein expression patterns allows direct investigation of DNA sequence translation. Therefore, methods for determining molecular mass composition are as valuable as sequencing of multiple loci.
Reducing the reproducibility of complex microbial communities based on mass spectrometric expression patterns provides new capabilities for scientific research (eg in environmental research and biodiversity surveys). Thousands of microbes can be easily analyzed by BioTyper and can then be used as a basis for further analysis. The initial microbiological reference spectrum atlas can be used to compare with each other and redundant can be removed. Further classified microorganisms can be compared to the initial reference data set and then removed or added to the spectral library to create an overview of the microbial polymorphism.