Electrospray ionization quadrupole time-of-flight mass spectrometry and quadrupole mass spectrometry for genotyping single nucleotide substitutions in intact polymerase chain reaction products in K-ras and p53

2002 ◽  
Vol 16 (24) ◽  
pp. 2278-2285 ◽  
Author(s):  
Warees T. Muhammad ◽  
Karen F. Fox ◽  
Alvin Fox ◽  
William Cotham ◽  
Michael Walla
Keyword(s):  
Mass Spectrometry ◽  
Polymerase Chain Reaction ◽  
Electrospray Ionization ◽  
Reaction Products ◽  
Quadrupole Mass ◽  
Chain Reaction ◽  
Nucleotide Substitutions ◽  
Single Nucleotide ◽  
Flight Mass Spectrometry ◽  
Polymerase Chain

scholarly journals Identification of Bacterial Plant Pathogens Using Multilocus Polymerase Chain Reaction/Electrospray Ionization-Mass Spectrometry

2008 ◽  
Vol 98 (11) ◽  
pp. 1156-1164 ◽  
Author(s):  
E. Postnikova ◽  
C. Baldwin ◽  
C. A. Whitehouse ◽  
A. Sechler ◽  
N. W. Schaad ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Polymerase Chain Reaction ◽  
Electrospray Ionization ◽  
Electrospray Ionization Mass Spectrometry ◽  
Ionization Mass Spectrometry ◽  
Distinct Advantage ◽  
Ionization Mass ◽  
Chain Reaction ◽  
Esi Ms ◽  
Polymerase Chain

Polymerase chain reaction/electrospray ionization-mass spectrometry (PCR/ESI-MS, previously known as “TIGER”) utilizes PCR with broad-range primers to amplify products from a wide array of organisms within a taxonomic group, followed by analysis of PCR amplicons using mass spectrometry. Computer analysis of precise masses allows for calculations of base compositions for the broad-range PCR products, which can then be compared to a database for identification. PCR/ESI-MS has the benefits of PCR in sensitivity and high-throughput capacity, but also has the distinct advantage of being able to detect and identify organisms with no prior characterization or sequence data. Existing broad range PCR primers, designed with an emphasis on human pathogens, were tested for their ability to amplify DNA of well characterized phytobacterial strains, as well as to populate the existing PCR/ESI-MS bacterial database with base counts. In a blinded panel study, PCR/ESI-MS successfully identified 93% of unknown bacterial DNAs to the genus level and 73% to the species/subspecies level. Additionally, PCR/ESI-MS was capable of detecting and identifying multiple bacteria within the same sample. The sensitivity of PCR/ESI-MS was consistent with other PCR based assays, and the specificity varied depending on the bacterial species. Preliminary tests with real life samples demonstrate a high potential for using PCR/ESI-MS systems for agricultural diagnostic applications.

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