Biochemistry, sometimes called biological chemistry, is the study of chemical processes within and relating to, living organisms.
EnZtek’s biochemical focus is on enzymes.
The science of enzymology goes back to the 1800’s. EnZtek has taken that history and perfected it into tests which provide faster results. The enzymatic mechanism consists of a unique substrate contained in the EnZtek test identifies the target enzyme produced by the target bacteria, and a match is made, resulting in a detectable reaction. This is depicted in the Lock and Key Analogy diagram.
Why do EnZtek tests provide bacteria detection faster than other tests available?
Because, EnZtek tests detect the enzymes produced by bacteria and a healthy bacteria can produce thousands of enzymes, especially during infections. Most other bacteria detection tests need to find an actual bacteria, which requires the bacteria to grow, and this takes time, usually 24 hours or more. Even for PCR tests, the bacteria must be grown before enough DNA can be obtained and replicated.
Known Enzymatic Facts:
Rate of Reaction
Enzymes typically react with between 1 and 10,000 molecules per second. Fast enzymes catalyze up to 500,000 molecules per second. Substrate concentration, enzyme concentration, types of media or broth used, temperature and pH affect the rate of enzyme reactions.
Also, RFU does not equal CFU
The numeric reading result on a fluorometer is a measurement of the fluorescent reaction of the enzyme and substrate molecules producing a numeric value in RFU or Relative Fluorescent Units. These enzyme molecules came from the bacteria present in the sample. CFU on the other hand stands for Colony Forming Unit which is the measurement used for traditional tests which are designed to count bacteria colonies. The EnZtek tests are Qualitative, not Quantitative, in other words, Presence/Absence, which is ideal for fast screening to see if the bacteria is present.
The graph below is an example of RFU results after 10 minutes of reaction time. The testing was conducted with culture media spiked with Total Organism Bacteria – beginning concentrations ~ 3 to 500 CFU/1ml of media.. First the short 5 hour incubation was performed at 37 degrees C. Next the test procedure were performed and after 10 minutes of reaction time, the fluorometer readings were taken. Pass <500 RFU Fail >501 RFU.
Table is an average of 3187 assays.
EnZtek’s tests are intended for real-world conditions where the enzymes are naturally found and can be detected quickly. Any EnZtek testing performed “off-label” such as in a laboratory will behave differently due to the “artificial” environment found in the laboratory setting which can affect the production of enzymes significantly.
It is not EnZtek's intent to replace any required testing. EnZtek's vision is to augment testing programs by providing an early screening tool to achieve these goals.
A Different Approach
Every industry has its own requirements and guidelines it must follow in regards to bacteria detection. EnZtek offers its tests to those who find value in conducting effective microbial surveillance, and can benefit from the early feedback EnZtek’s tests offer in regards to detecting the presence of bacteria. Below is one scenario which can be utilized when using the EnZquik Fluorescent Assays: