Heat-killed cells of each AFB isola

Heat-killed cells of each AFB isolate were prepared by
mixing *2 loopful of cells (20 ll cell pellet) in 200 ll
dH2O followed by incubation at 80C for 1 h. Heat-killed
AFB samples were used as templates in multiplex polymerase
chain reactions (PCR) for typing of Mycobacterium
genus and region of difference (RD; deletion typing),
according to protocols previously described (Berg et al.
2009). Each isolate characterized as non-tuberculous
mycobacteria (NTM) was sequenced at the 16S rDNA locus
and the sequence was entered in the Basic Local Assignment
Search Tool (BLAST) database at the National Center
for Biotechnology Information (NCBI) and the Ribosomal
Differentiation of Microorganisms (RIDOM) (http://rdna.
ridom.de) database for further identification of species
(Berg et al. 2009). DNA sequencing was performed at
the Animal Health and Veterinary Laboratories Agency
(AHVLA), United Kingdom, using an Applied Biosystems
model 3730 automated capillary DNA sequencer. Isolates
genetically identified by deletion typing as of the M. tuberculosis
complex (MTC) were spoligotyped for further strain
characterization as previously described (Kamerbeek et al.
1997). Spoligotyping data were compared with the Spoligo-
International-Typing (SIT) database (http://www.pasteurguadeloupe.
fr:8081/SITVITDemo/ and http://www.cs.rpi.
edu/*bennek/tbinsight/tblineage.html to match SIT numbers
and lineage classifications. Isolates identified as
M. bovis were compared with spoligotype patterns in the
international M. bovis database (www.mbovis.org). Spoligotype
patterns of all MTC isolates were analysed using
spolTools (http://www.emi.unsw.edu.au/spolTools) (Tang
et al. 2008).

Hasil (Bahasa Indonesia) 1:

Heat-killed cells of each AFB isolate were prepared bymixing *2 loopful of cells (20 ll cell pellet) in 200 lldH2O followed by incubation at 80C for 1 h. Heat-killedAFB samples were used as templates in multiplex polymerasechain reactions (PCR) for typing of Mycobacteriumgenus and region of difference (RD; deletion typing),according to protocols previously described (Berg et al.2009). Each isolate characterized as non-tuberculousmycobacteria (NTM) was sequenced at the 16S rDNA locusand the sequence was entered in the Basic Local AssignmentSearch Tool (BLAST) database at the National Centerfor Biotechnology Information (NCBI) and the RibosomalDifferentiation of Microorganisms (RIDOM) (http://rdna.ridom.de) database for further identification of species(Berg et al. 2009). DNA sequencing was performed atthe Animal Health and Veterinary Laboratories Agency(AHVLA), United Kingdom, using an Applied Biosystemsmodel 3730 automated capillary DNA sequencer. Isolatesgenetically identified by deletion typing as of the M. tuberculosiscomplex (MTC) were spoligotyped for further straincharacterization as previously described (Kamerbeek et al.1997). Spoligotyping data were compared with the Spoligo-International-Typing (SIT) database (http://www.pasteurguadeloupe.fr:8081/SITVITDemo/ and http://www.cs.rpi.edu/*bennek/tbinsight/tblineage.html to match SIT numbersand lineage classifications. Isolates identified asM. bovis dibandingkan dengan pola spoligotype diInternasional M. bovis database (di www.mbovis.org). Spoligotypepola semua isolat MTC dianalisa dengan menggunakanspolTools (Tang (http://www.emi.unsw.edu.au/spolTools)et al., 2008).