title: Loop-mediated isothermal amplification (LAMP) test for specific and rapid detection of Brucella abortus in cattle.
abstract: BACKGROUND
  Brucella abortus, the major causative agent of abortion in cattle and a zoonotic pathogen, needs to be diagnosed at an early stage. Loop-mediated isothermal amplification (LAMP) test is easy to perform and also promising to be adapted at field level.
  
  
  OBJECTIVE
  To develop a LAMP assay for specific and rapid detection of B. abortus from clinical samples of cattle.
  
  
  METHODS
  LAMP primers were designed targeting BruAb2_0168 region using specific software tool and LAMP was optimized. The developed LAMP was tested for its specificity with 3 Brucella spp. and 11 other non-Brucella spp. Sensitivity of the developed LAMP was also carried out with known quantity of DNA. Cattle whole blood samples and aborted fetal stomach contents were collected and used for testing with developed LAMP assay and results were compared with polymerase chain reaction (PCR).
  
  
  RESULTS
  The developed LAMP assay works at 61 °C for 60 min and the detection limit was observed to be 100-fold more than the conventional PCR that is commonly used for diagnosis of B. abortus. Clinical sensitivity and specificity of the developed LAMP assay was 100% when compared with Rose Bengal plate test and standard tube agglutination test. SYB® green dye I was used to visualize the result with naked eye.
  
  
  CONCLUSIONS
  The novelty of the developed LAMP assay for specifically detecting B. abortus infection in cattle along with its inherent rapidness and high sensitivity can be employed for detecting this economically important pathogen of cattle at field level as well be exploited for screening of human infections.
authors.affiliation: a Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Bareilly, India.
authors.forename: K
authors.initials: K
authors.lastname: Karthik
authors.forename: Rajesh
authors.initials: R
authors.lastname: Rathore
authors.forename: Prasad
authors.initials: P
authors.lastname: Thomas
authors.forename: T R
authors.initials: TR
authors.lastname: Arun
authors.forename: K N
authors.initials: KN
authors.lastname: Viswas
authors.forename: R K
authors.initials: RK
authors.lastname: Agarwal
authors.forename: H V
authors.initials: HV
authors.lastname: Manjunathachar
authors.forename: Kuldeep
authors.initials: K
authors.lastname: Dhama
mesh_terms: D000818:Animals; D002002:Brucella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D002003:Brucella abortus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002007:Brucellosis, Bovine / Q000175:diagnosis* / Q000382:microbiology; D002417:Cattle; D005260:Female; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary*; D012680:Sensitivity and Specificity
keywords: BruAb2_0168; Brucella abortus; LAMP; cattle; cow; whole blood