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Enterotoxigenic Escherichia coli is detectable in water samples from an endemic area by real-time PCR.

Artikel i vetenskaplig tidskrift
Författare Åsa Lothigius
Anders Janzon
Y Begum
Åsa Sjöling
F Qadri
Ann-Mari Svennerholm
Ingrid Bölin
Publicerad i Journal of applied microbiology
Volym 104
Nummer/häfte 4
Sidor 1128-36
ISSN 1365-2672
Publiceringsår 2008
Publicerad vid Institutionen för biomedicin, avdelningen för mikrobiologi och immunologi
Sidor 1128-36
Språk en
Länkar dx.doi.org/10.1111/j.1365-2672.2007...
Ämnesord Bacterial Toxins, genetics, Bangladesh, Colony Count, Microbial, DNA Primers, genetics, DNA, Bacterial, analysis, Enterotoxigenic Escherichia coli, genetics, Enterotoxins, genetics, Environmental Monitoring, methods, Enzyme-Linked Immunosorbent Assay, methods, Escherichia coli Proteins, genetics, Reverse Transcriptase Polymerase Chain Reaction, methods, Water Microbiology, Water Supply
Ämneskategorier Mikrobiologi inom det medicinska området

Sammanfattning

AIMS: We aimed to develop an assay for sensitive detection and quantification of enterotoxigenic Escherichia coli (ETEC) in different types of water samples. METHODS AND RESULTS: Real-time polymerase chain reaction (PCR) assays with primers against ETEC enterotoxin genes estA (STh) estB (STp) and eltB (LT) were designed and the detection levels were determined to be three bacteria per PCR reaction. Gene copy numbers were estimated to be four (LT), two (STh) and one (STp) per bacteria. Twenty-six household and 13 environmental water samples from Bangladesh were filtered through 0.22-microm filters; DNA was extracted from the filters and analysed by real-time PCR. The results were compared with toxin GM1-enzyme-linked immunosorbent assay (ELISA), in which colonies were tested for toxin production after cultivation of the filters. Out of the 39 samples tested, 18 household and 8 environmental samples were positive for ETEC in real-time PCR, but only 6 positive samples were found with GM1-ELISA. CONCLUSIONS: The method allows for highly sensitive detection and quantification of ETEC based on detection of toxin DNA in water samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The method facilitates detection and identification of ETEC in water and allows comparison between water contamination and incidence of ETEC diarrhoea in endemic areas.

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