PMID- 22747670
OWN - NLM
STAT- MEDLINE
DCOM- 20121217
LR  - 20211021
IS  - 1746-6148 (Electronic)
IS  - 1746-6148 (Linking)
VI  - 8
DP  - 2012 Jun 29
TI  - Isolation of Mycobacterium avium subspecies paratuberculosis from Ugandan cattle 
      and strain differentiation using optimised DNA typing techniques.
PG  - 99
LID - 10.1186/1746-6148-8-99 [doi]
AB  - BACKGROUND: The occurrence of paratuberculosis in Ugandan cattle has recently 
      been reported but there is no information on the strains of Mycobacterium avium 
      subspecies paratuberculosis (MAP) responsible for the disease. The aim of this 
      study was to isolate and characterise MAP from seropositive cattle and 
      paratuberculosis lesions in tissues obtained from slaughtered cattle in Uganda. 
      RESULTS: Twenty one isolates of MAP were differentiated into 11 genotype profiles 
      using seven genotyping loci consisting of Insertion Sequence 1311(IS1311), 
      Mycobacterial interspersed repeat units (MIRU) (loci 2, 3), Variable number 
      tandem repeats (VNTR) locus 32 and Short sequence repeats (SSR) (loci 1, 2 and 
      8). Three different IS1311 types and three MIRU 2 profiles (7, 9, 15 repeats) 
      were observed. Two allelic variants were found based on MIRU 3 (1, 5 repeats), 
      while VNTR 32 showed no polymorphism in any of the isolates from which it was 
      successfully amplified. SSR Locus 1 revealed 6 and 7 G1 repeats among the 
      isolates whereas SSR locus 2 revealed 10, 11 and 12 G2 repeats. SSR locus 8 was 
      the most polymorphic locus. Phylogenetic analysis of SSR locus 8 sequences based 
      on their single nucleotide polymorphisms separated the isolates into 8 genotypes. 
      We found that the use of Ethylene glycol as a PCR additive improved the 
      efficiency of the PCR reactions for MIRUs (2, 3), VNTR 32 and SSR (loci 1 and 2). 
      CONCLUSIONS: There is a high strain diversity of MAP in Uganda since 21 isolates 
      could be classified into 11 genotypes. The combination of the seven loci used in 
      this study results into a very precise discrimination of isolates. However 
      analysis of SNPs on locus alone 8 is very close to this combination. Most of the 
      genotypes in this study are novel since they differed in one or more loci from 
      other isolates of cattle origin in different studies. The large number of MAP 
      strains within a relatively small area of the country implies that the 
      epidemiology of paratuberculosis in Uganda may be complicated and needs further 
      investigation. Finally, the use of Ethylene glycol as a PCR additive increases 
      the efficiency of PCR amplification of difficult templates.
FAU - Okuni, Julius Boniface
AU  - Okuni JB
AD  - College of Veterinary Medicine, Animal Resources and Biosecurity, Makerere 
      University, P.O. Box 7062, Kampala, Uganda. jbok@vetmed.mak.ac.ug
FAU - Dovas, Chrysostomos I
AU  - Dovas CI
FAU - Loukopoulos, Panayiotis
AU  - Loukopoulos P
FAU - Bouzalas, Ilias G
AU  - Bouzalas IG
FAU - Kateete, David Patrick
AU  - Kateete DP
FAU - Joloba, Moses L
AU  - Joloba ML
FAU - Ojok, Lonzy
AU  - Ojok L
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120629
PL  - England
TA  - BMC Vet Res
JT  - BMC veterinary research
JID - 101249759
RN  - 0 (DNA, Bacterial)
SB  - IM
MH  - Animals
MH  - Bacteriological Techniques/*veterinary
MH  - Cattle
MH  - DNA Fingerprinting/*methods
MH  - DNA, Bacterial/classification/genetics
MH  - Genetic Variation
MH  - Genotype
MH  - Mycobacterium avium subsp. paratuberculosis/classification/*genetics
MH  - Paratuberculosis/epidemiology/*microbiology
MH  - Phylogeny
MH  - Polymorphism, Restriction Fragment Length
MH  - Uganda/epidemiology
PMC - PMC3416654
EDAT- 2012/07/04 06:00
MHDA- 2012/12/18 06:00
PMCR- 2012/06/29
CRDT- 2012/07/04 06:00
PHST- 2012/02/24 00:00 [received]
PHST- 2012/06/18 00:00 [accepted]
PHST- 2012/07/04 06:00 [entrez]
PHST- 2012/07/04 06:00 [pubmed]
PHST- 2012/12/18 06:00 [medline]
PHST- 2012/06/29 00:00 [pmc-release]
AID - 1746-6148-8-99 [pii]
AID - 10.1186/1746-6148-8-99 [doi]
PST - epublish
SO  - BMC Vet Res. 2012 Jun 29;8:99. doi: 10.1186/1746-6148-8-99.