title: Isolation & characterization of Brucella melitensis isolated from patients suspected for human brucellosis in India.
abstract: OBJECTIVE
  Brucellosis is endemic in the southern part of India. A combination of biochemical, serological and molecular methods is required for identification and biotyping of Brucella. The present study describes the isolation and biochemical, molecular characterization of Brucella melitensis from patients suspected for human brucellosis.
  
  
  METHODS
  The blood samples were collected from febrile patients suspected to have brucellosis. A total of 18 isolates were obtained from 102 blood samples subjected to culture. The characterization of these 18 isolates was done by growth on Brucella specific medium, biochemical reactions, CO2 requirement, H2S production, agglutination with A and M mono-specific antiserum, dye sensitivity to basic fuchsin and thionin. Further, molecular characterization of the isolates was done by amplification of B. melitensis species specific IS 711 repetitive DNA fragment and 16S (rRNA) sequence analysis. PCR-restriction fragment length polymorphism (RFLP) analysis of omp2 locus and IS711 gene was also done for molecular characterization.
  
  
  RESULTS
  All 102 suspected samples were subjected to bacteria isolation and of these, 18 isolates could be recovered on blood culture. The biochemical, PCR and PCR-RFLP and 16s rRNA sequencing revealed that all isolates were of B. melitensis and matched exactly with reference strain B. melitensis 16M.
  
  
  CONCLUSIONS
  The present study showed an overall isolation rate of 17.64 per cent for B. melitensis. There is a need to establish facilities for isolation and characterization of Brucella species for effective clinical management of the disease among patients as well as surveillance and control of infection in domestic animals. Further studies are needed from different geographical areas of the country with different level of endemicity to plan and execute control strategies against human brucellosis.
authors.affiliation: Microbiology Division, Defence Research and Development Establishment, Gwalior, Bijapur, India.
authors.forename: Anita
authors.initials: A
authors.lastname: Barua
authors.affiliation: Microbiology Division, Defence Research and Development Establishment, Gwalior, Bijapur, India.
authors.forename: Ashu
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Microbiology Division, Defence Research and Development Establishment, Gwalior, Bijapur, India.
authors.forename: Duraipandian
authors.initials: D
authors.lastname: Thavaselvam
authors.affiliation: Microbiology Division, Shri BM Patil Medical College, BLDE University, Bijapur, India.
authors.forename: Smita
authors.initials: S
authors.lastname: Mangalgi
authors.affiliation: Microbiology Division, Defence Research and Development Establishment, Gwalior, Bijapur, India.
authors.forename: Archana
authors.initials: A
authors.lastname: Prakash
authors.affiliation: Microbiology Division, Defence Research and Development Establishment, Gwalior, Bijapur, India.
authors.forename: Sapana
authors.initials: S
authors.lastname: Tiwari
authors.affiliation: Microbiology Division, Defence Research and Development Establishment, Gwalior, Bijapur, India.
authors.forename: Sonia
authors.initials: S
authors.lastname: Arora
authors.affiliation: Microbiology Division, Defence Research and Development Establishment, Gwalior, Bijapur, India.
authors.forename: Kannusamy
authors.initials: K
authors.lastname: Sathyaseelan
mesh_terms: D001483:Base Sequence / Q000235:genetics; D017347:Brucella melitensis / Q000302:isolation & purification* / Q000472:pathogenicity; D002006:Brucellosis / Q000097:blood* / Q000382:microbiology / Q000473:pathology; D006801:Humans; D007194:India; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics*
chemical_list: D012336:RNA, Ribosomal, 16S