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Detection of low numbers of pathogenic Yersinia enterocolitica in environmental water and sewage samples by nested polymerase chain reaction
Author(s) -
Waage A. S.,
Vardund T.,
Lund V.,
Kapperud G.
Publication year - 1999
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.1999.00915.x
Subject(s) - yersinia enterocolitica , nested polymerase chain reaction , polymerase chain reaction , sewage , microbiology and biotechnology , biology , enteric virus , bacteria , gene , genetics , environmental science , environmental engineering
A.S. WAAGE, T. VARDUND, V. LUND and G. KAPPERUD.1999.Isolation of pathogenic Yersinia enterocolitica from water and sewage by traditional culture techniques is time‐consuming and subsequent differentiation between pathogenic and non‐pathogenic strains can be difficult and unreliable. A nested polymerase chain reaction (PCR) procedure was used for the detection of low numbers of Y. enterocolitica in spiked samples from natural surface sources with variable background flora ranging from oligotrophic water to sewage. Water and sewage samples were filtered and filters enriched overnight in a non‐selective medium. Nested PCR conducted on enriched broth, prepared by use of a rapid and simple preparation step consisting of centrifugation, proteinase K treatment and boiling, enabled the detection of 8–17 cfu 100 ml −1 water with background levels of up to 8700 heterotrophic organisms ml −1 and 10 000 cfu coliform organisms 100 ml −1 water. The analysis can be completed within 2–3 d and should be a significant tool in monitoring environmental waters and drinking water sources for the presence of pathogenic Y. enterocolitica .