BIOCHEMICAL DETECTION METHODS FOR BACTERIA AND VIRUSES
Quarterly progress rept. no. 3, 1 Jan-31 Mar 1963
MELPAR FALLS CHURCH VA
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Various factors affecting the bacterial este-3were studied and evaluated by means of a stand ardized microphotofluorimeter. Among these were heat denaturation, comparison of other substrates, and salt effects. Thirteen different species of bacteria were examined for esterases to hydrolyze fluorescein diacetate. The enzymes responsible for hydrolysis of phenylacetate and FDA are both universally present in all bacteria, including the anaerobes which were not previously tested. The amplified enzyme approach for detection was tested using papain. Various activators, in cluding whole and broken bacterial cells, were examined for their ability to activate papain. Activation by bacteria required large numbers of cells, and possible methods of amplifying these effects were explored. A system containing cholinesterase, acetylthiocholine, papain, and CTN was used to demonstrate that the product from one enzyme system can activate a second enzyme. Various methods for applying this approach to detection of bacteria are suggested.