Lysin Agent and Method of Use for Diagnostic Testing

Abstract

Novel Bacillus lysin proteins have been identified and characterized. Methods to detect the presence or absence of bacteria in a sample, methods for lysing bacteria, and methods for decontaminating or disinfecting areas contaminated with bacteria that utilize these lysin proteins have also been developed.

What is claimed: 
     
         1 . A method for detecting the presence or absence of bacteria in a sample comprising:
 incubating the sample in the presence of at least one polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12;   contacting the sample with at least a first and a second oligonucleotide primer under conditions sufficient to provide polymerase-based nucleic acid amplification; and   detecting the presence or absence of an amplification product.   
     
     
         2 . The method of  claim 1 , wherein the bacteria is a Gram-positive bacteria. 
     
     
         3 . The method of  claim 1 , wherein the bacteria is a  Bacillus  species. 
     
     
         4 . The method of  claim 1 , wherein the bacteria is  Bacillus anthracis.    
     
     
         5 . The method of  claim 1 , wherein the bacteria is a Gram-negative bacteria. 
     
     
         6 . The method of  claim 1 , wherein the bacteria is a  Yersinia  species. 
     
     
         7 . The method of  claim 1 , wherein the bacteria is  Yersinia pestis.    
     
     
         8 . The method of  claim 1 , wherein the polymerase-based nucleic acid amplification is quantitative polymerase chain reaction (QPCR). 
     
     
         9 . The method of  claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 10. 
     
     
         10 . A method for lysing bacteria, comprising contacting the bacteria with at least one polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12. 
     
     
         11 . The method of  claim 10 , wherein the bacteria is a Gram-positive bacteria. 
     
     
         12 . The method of  claim 10 , wherein the bacteria is a  Bacillus  species. 
     
     
         13 . The method of  claim 10 , wherein the bacteria is  Bacillus anthracis.    
     
     
         14 . The method of  claim 10 , wherein the bacteria is a Gram-negative bacteria. 
     
     
         15 . The method of  claim 10 , wherein the bacteria is a  Yersinia  species. 
     
     
         16 . The method of  claim 10 , wherein the bacteria is  Yersinia pestis.    
     
     
         17 . The method of  claim 1 , wherein the at least one polypeptide is in the form of a powder. 
     
     
         18 . The method of  claim 10 , wherein the at least one polypeptide is in the form of a powder. 
     
     
         19 . The method of  claim 1 , wherein the at least one polypeptide further comprises a tag. 
     
     
         20 . The method of  claim 10 , wherein the at least one polypeptide further comprises a tag. 
     
     
         21 . A method for decontaminating or disinfecting a surface or room or area or object contaminated with bacteria, comprising contacting the surface or room or area or object with a composition comprising at least one polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO:  12 . 
     
     
         22 . The method of  claim 21 , wherein the composition is in the form of a powder.