Method for the detection of bacterial species of the genera anaplasma/ehrlichia and bartonella

Abstract

The present invention relates to a method for the detection and identification of bacterial species belonging to the genera Anaplasma/Ehrlichia and Bartonella , and also provides triggers and probes required for its application, as well as associated kits.

1 . A method for detecting bacterial species that cause zoonosis, belonging to the genera  Anaplasma/Ehrlichia  and  Bartonella , comprising the following steps:
 a. placing the sample under analysis in contact with a reaction mixture containing specific triggers that will amplify one or more sequences selected from the group comprising SEQ ID NO:1-17 and 47 or their respective complementary sequences;   b. amplifying the sequences by means of polymerase chain reaction; and   c. detecting amplification products formed in the previous step, said detection being indicative of the presence or absence of zoonosis-causing bacteria.   
     
     
         2 . The method according to  claim 1 , further comprising the step of identifying the bacterial species corresponding to the amplification products. 
     
     
         3 . The method of  claim 2 , wherein the steps of detecting and identifying are simultaneous. 
     
     
         4 . The method according to  claim 1 , wherein the triggers comprise sequences selected from SEQ ID NO:18-25 and their complementary sequences. 
     
     
         5 . The method according to  claim 1 , in which the detection of amplification products comprises using one or more probes having a sequence selected from SEQ ID NO:26-42 and 48 and their complementary sequences. 
     
     
         6 . A trigger having a sequence selected from SEQ ID NO:20-21 and their complementary sequences, said trigger allowing the amplification of a sequence selected from SEQ ID NO:2-7 and 47. 
     
     
         7 . The trigger of  claim 6 , wherein the trigger has a sequence selected from SEQ ID NO:22-23 and their complementary sequences, said trigger allowing the amplification of SEQ ID NO:8. 
     
     
         8 . A probe having a sequence selected from SEQ ID NO:26-42 and 48 and their complementary sequences, said probe being capable of hybridizing with a sequence selected from SEQ ID NO:1-17 and 47 and their complementary sequences. 
     
     
         9 . A kit for detecting bacterial species that cause zoonosis, belonging to the genera  Anaplasma/Ehrlichia  and  Bartonella , comprising one or more triggers having a sequence selected from SEQ ID NO:18-25 and their complementary sequences. 
     
     
         10 . The kit according to  claim 9 , further comprising one or more probes having a sequence selected from SEQ ID NO:26-42 and 48 and their complementary sequences.