By Stephen H. Gillespie
Scientific and diagnostic microbiologists element molecular and actual tools for learning the turning out to be challenge of antibiotic resistance in micro organism, and facilitate new antibiotic examine courses to aid redress the matter. The options variety from those who offer swift analysis via DNA amplifications and phage reveal, to these for plotting the transmission of resistant organisms and investigating their epidemiology. in addition to illuminating the fundamental biology of antimicrobial resistance, in addition they strengthen and enforce now diagnostic and epidemiological instruments.
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Additional info for Antibiotic Resistance: Methods and Protocols
24. 25. 26. 27. Incubator, 37°C. Colorimeter (Viteck). Exhaust protective biological safety cabinet (NuAire, Plymounth, MN). Mycobacterium tuberculosis (Mtb), antibiotic sensitive strain H37Rv ATCC 27294 (American Type Culture Collection). Mtb, rifampin resistant strain ATCC 35838 (American Type Culture Collection). Mtb, isoniazid resistant strain, CAP E-05, 1995, Set E-A (College of American Pathology). Middlebrook 7H9 broth. Lowenstein-Jensen slants. BBL® MycoPrep™ Specimen Digestion/Decontamination Kit (Becton Dickinson, Sparks, MD).
For instance, the multi-antibiotic sensitive strain, H37Rv in the presence of isoniazid or rifampin, had a negative slope whose 95% confidence interval did not overlap the confidence interval of the 1% control (Fig. 6). ). Similarly, the isoniazid resistant strain, CAP E-05, had a mildly positive growth slope and its 95% confidence interval did not overlap that of the 1% control (Fig. 5A). However, QSTN-PCR for M. tuberculosis Susceptibility Testing 51 Fig. 5. Growth slope analysis of resistant organisms.
3. Aliquot the master mix into thin walled PCR reaction tubes (see Note 7). 5. Add an aliquot of 10 µL template DNA (sample, genomic DNA or sterile distilled water) to each tube and mixed (see Note 8). 6. Overlay the reaction mix with 3 drops (approx 50 µL) mineral oil. 7. Transfer the reaction tubes to a thermal cycler and perform the amplification under the following conditions: one cycle of 95°C for 1 min; thirty cycles of 94°C for 1 min, 65°C for 2 min, 72°C for 3 min; one cycle of 72°C for 7 min.
Antibiotic Resistance: Methods and Protocols by Stephen H. Gillespie