US2013237457A1PendingUtilityA1

Method of detecting pathogenic legionella strains

Assignee: NATUURWETENSCHAPPELIJK ONDERZOEK TNO NL ORGANISATIE VOOR TOEGEPASTPriority: Dec 29, 2006Filed: Apr 3, 2013Published: Sep 12, 2013

Est. expiryDec 29, 2026(~0.4 yrs left)· nominal 20-yr term from priority

Inventors:Frank Henri Johan Schuren Roy Christiaan Montijn Henricus Matheus Wilhelmus Maria Thijssen

Y02A50/30C12Q 1/689

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References

Claims

Abstract

The invention relates to a kit for detecting pathogenic Legionella pneumophila strains by hybridizing genomic DNA of a sample suspected to contain Legionella to two or five specific sequence markers, as identified by MARKER NO. 1 through MARKER NO. 5 or homologues thereof.

Claims

exact text as granted — not AI-modified

1 . A hybridization assay kit for use in a method to determine whether a  Legionella pneumophila  strain is a pathogenic strain or a non-pathogenic strain in a sample containing  Legionella pneumophila,  which comprises an array of less than 50 nucleotide sequences and which comprises a nucleotide sequence comprising at least 50 contiguous nucleotides from MARKER NO. 1 and a nucleotide sequence comprising at least 50 contiguous nucleotides from MARKER NO. 2 or homologues thereof. 
     
     
         2 . A hybridization assay kit according to  claim 1 , which further comprises nucleotide sequences comprising at least 50 contiguous nucleotides from each of MARKER NO. 3, MARKER NO. 4 and MARKER NO. 5 or homologues thereof. 
     
     
         3 . A hybridization assay kit according to  claim 1 , which further comprises a positive control sequence. 
     
     
         4 . A hybridization assay kit according to  claim 1 , which further comprises software for calculating the results of the logic rule
 If (MARKER NO. 1&gt;MARKER NO. 2) then said strain is pathogenic   wherein MARKER NO. 1 and MARKER NO. 2 are stochastic values representing 1 if hybridization and 0 if no hybridization has been observed with said sequences.   
     
     
         5 . A hybridization assay kit according to  claim 2 , which further comprises software for calculating the results of the logic rule
 (1) If (MARKER NO. 2&lt;MARKER NO. 1) then [not(MARKER NO. 2=MARKER NO. 3)] else [(MARKER NO. 2=MARKER NO. 4) and (MARKER NO. 5=MARKER NO. 3)]   (2) If (MARKER NO. 2&lt;MARKER NO. 3) then [not(MARKER NO. 1&lt;MARKER NO. 4)] else [(MARKER NO. 5&gt;MARKER NO. 2) nor (MARKER NO. 5&lt;MARKER NO. 4)](3) If (MARKER NO. 2&lt;MARKER NO. 3) then [not(MARKER NO. 4&lt;MARKER NO. 1)] else [(MARKER NO. 3&lt;MARKER NO. 5) nor (MARKER NO. 4&gt;MARKER NO. 5)]   (4) If (MARKER NO. 1=MARKER NO. 5) then [(MARKER NO. 4&lt;=MARKER NO. 1)] else [(MARKER NO. 1&lt;MARKER NO. 4) nor (MARKER NO. 3=MARKER NO. 2)]   (5) If (MARKER NO. 1=MARKER NO. 5) then [ (MARKER NO. 5&gt;=MARKER NO. 4)] else [(MARKER NO. 3&lt;=MARKER NO. 2) nor (not(MARKER NO. 4&lt;MARKER NO. 1))]   (6) If (MARKER NO. 1=MARKER NO. 5) then [not(MARKER NO. 4&gt;MARKER NO. 1)] else [(MARKER NO. 4&gt;MARKER NO. 1) nor (MARKER NO. 2&gt;=MARKER NO. 3)]   (7) If (MARKER NO. 1=MARKER NO. 5) then [not(MARKER NO. 1&lt;MARKER NO. 4)] else [(MARKER NO. 3=MARKER NO. 2) nor (MARKER NO. 1&lt;MARKER NO. 4)],   wherein MARKER NO. n (n=1-5) is the stochastic value representing 1 if hybridization and 0 if no hybridization has been observed with said sequence n, and wherein said strain is pathogenic if 4 or more of the logic rules (1)-(7) have the result TRUE.   
     
     
         6 . A hybridization assay kit according to  claim 3 , wherein the positive control sequence is  Legionella  30S ribosomal protein S21 or a conserved sequence of one of the  Legionella  housekeeping enzymes.

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