Page 52 - Тахианаас хүнсний хордлого үүсгэгч CAMPYLOBACTER SPP.-ийг илрүүлсэн судалгааны дүн

[ШИНЖЛЭХ УХААНЫ МАГИСТРЫН ЗЭРЭГ ГОРИЛСОН БҮТЭЭЛ] Хураангуй
Дүгнэлт

The strains were cultured on Mueller–Hinton agar containing 5% sheep blood under
microaerobic conditions (N2: 85%, O2:5%,CO2: 10%) at 378C for 48 h. The minimal inhibitory
concentrations (MICs) of erythromycin and ciprofloxacin were determined in duplicate by agar
dilution method according to the CLSI recommendations (Clinical and Laboratory Standards

Institute, 2002). The resistance breakpoints were ciprofloxacin, ofloxacin, tetracycline, naladix
acid, norflaxacin.
DNA extraction, PCR and sequencing analysis
Sequence analysis showed that the macrolide-resistant isolates had a point mutation in the 23S
ribosomal RNA (rRNA) genes (A2075G) and that the fluoroquinolone-resistant isolates had a
point mutation in the gyrase gene gyrA (C257T
Genomic DNA was extracted using Puregene DNA Purification Kit (Gentra Systems, Inc.,
Minneapolis,MN). The 285-bp fragment covering the quinolone resistance-determining region of
gyrA from C. Coli and C.jejuni was amplified by PCR using the forward primer gyrA-campy-F:5’-
GAGTGTTATTATAGGTCGTGC-3’ and the reverse primer gyrA-campy-R:5’-
GGCACTATCACCATCTATAG-3’, Sequencing was achieved on a Prism 3130xl (Applied
biosystems, USA ) Genetic Analyzer using dRhodamine-labeled terminators.

The following seven loci were chosen for the MLST scheme (protein products are shown in
parentheses): aspA(aspartase A), glnA(glutamine synthetase), gltA(citrate synthase),
glyA(serine hydroxymethyl-transferase), pgm(phosphoglucomutase), tkt (transketolase), and
uncA (ATP synthase asubunit). The chromosomal locations of these housekeeping loci
suggested that it was unlikely for any of them to be coinherited in the same recombination event,
as the minimum distance between loci was 70 kb.

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