Realtids-PCR för påvisande av plasmidburen ampicillinresistens: Kartläggning av förekomst i vattenisolat från Helge Å, Kristianstad
2018 (Swedish) Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE credits
Student thesis Alternative title
Real-Time PCR for detection of plasmid-mediated ampicillinresistance : A survey of prevalence in water isolates from Helge river, Kristianstad (English)
Abstract [en]
The antibiotic class β-lactams include drugs such as penicillins, cephalosporines, carbapenems and monobactams which mechanism of action is to inhibit cell-wall synthesis. Bacteria have developed several mechanisms to counter β-lactams. Bacteria can defend themselves from antibiotics by releasing enzymes that attack the antibiotic compound itself by hydrolysis, target alteration or redox reactions. Presence of antibiotics can also trigger a downregulation of genes coding for antibiotic binding proteins, as well as upregulation of proteins that serves as channel and pump proteins that ensure no accumulation of antibiotics occurs in the cytosol. The aim with the study was to investigate the presence of three plasmid-mediated genes (bla FOX, bla CIT(CMY-2) and bla MOX) coding for ampicillin resistance (pAmpC) in water isolates sampled from Helge River, Kristianstad. The detection of genes was done according to a previous optimized protocol for Real-Time PCR with SYBR™Green chemistry (duplex bla CIT(CMY-2)/bla MOX and singleplex bla FOX). The method proved not to be robust for multiplex PCR, only the singelplex for the gene bla FOX could produce valid results. 30 of 96 isolates were deemed as positive for the gene, whereas 27 of 79 were considered clinical relevant. Among the 27 isolates, 16 also harbored other genes for resistance (13 bla CTX-M, 2 bla OXA, 1 bla TEM and 1 bla SHV). One isolate carried on three resistancegenes (bla FOX, bla CTX-M och bla TEM). A majority of the positive isolates, 20 out of 27, were sampled near the pumpstation. The findings indicate that Helge river might be a reservoir for dissemination of antibiotic resistance genes.
Place, publisher, year, edition, pages 2018. , p. 43
Keywords [sv]
Ampicillinresistens, Realtids-PCR, AmpC
National Category
Biomedical Laboratory Science/Technology
Identifiers URN: urn:nbn:se:hkr:diva-18831 OAI: oai:DiVA.org:hkr-18831 DiVA, id: diva2:1263911
Subject / course Biomedical Laboratory Science
Educational program Bachelor of Science in Biomedical Laboratory Science
Supervisors
Examiners
2018-11-212018-11-182018-11-21 Bibliographically approved