title: Evaluation of complementary diagnostic tools for bovine tuberculosis detection in dairy herds from India.
abstract: OBJECTIVE
  A cross-sectional study was undertaken to know the herd prevalence and evaluate the single intradermal tuberculin testing (SITT), culture isolation, and polymerase chain reaction (PCR) analysis for the diagnosis of bovine tuberculosis (TB).
  
  
  METHODS
  A total of 541 cows of three dairy farms of Bareilly and Mukteshwar were screened by SITT followed by collection of pre-scapular lymph node (PSLN) aspirates (71), milk (54), and blood (71) samples from reactor animals. These clinical samples were processed for culture isolation and direct PCR-based identification and species differentiation.
  
  
  RESULTS
  Out of 541 cows screened by SITT, 71 (13.12%) animals were found positive. Mycobacteria were isolated from 3 (4.22%) PSLN aspirate but not from any cultured milk and blood samples. 28 (39.43%) PSLN aspirate and 5 (9.25%) milk samples were positive for Mycobacterium TB (MTB) complex (MTC) by PCR amplification for the IS6110 insertion sequence; however, blood samples were found negative. For species differentiation, multiplex-PCR using 12.7 kb primers was conducted. Out of 28 PSLN aspirate, Mycobacterium bovis was detected in 18 (64.28%) and MTB in 8 (28.57%), whereas 2 aspirate samples (7.14%) were positive for both the species. All the five milk positive samples were positive for M. bovis.
  
  
  CONCLUSIONS
  Direct detection of bovine TB by a molecular-based method in dairy animals after preliminary screening was appeared to be more sensitive and specific compared to the conventional method (i.e., culture isolation). Its application in form of serial testing methodology for the routine diagnosis and thereafter, culling of infected stock may be suggested for the control programs in dairy herds. The PSLN aspirate was found to be the most suitable specimen for culture isolation and PCR-based detection of Mycobacterium spp. among live infected animals.
authors.affiliation: School of Public Health & Zoonoses, Guru Angad Dev Veterinary & Animal Sciences University, Ludhiana, Punjab, India.
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Thakur
authors.affiliation: Division of Epidemiology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India.
authors.forename: Dharmender Kumar
authors.initials: DK
authors.lastname: Sinha
authors.affiliation: Division of Epidemiology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India.
authors.forename: Bhoj Raj
authors.initials: BR
authors.lastname: Singh
keywords: Mycobacterium tuberculosis complex; bovine tuberculosis; culture isolation; polymerase chain reaction; single intradermal tuberculin testing