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Detection of Escherichia coli O157:H7 in soil and water using multiplex PCR
Author(s) -
Campbell G.R.,
Prosser J.,
Glover A.,
Killham K.
Publication year - 2001
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.1365-2672.2001.01465.x
Subject(s) - escherichia coli , multiplex , multiplex polymerase chain reaction , pathogen , inoculation , biology , microbiology and biotechnology , incubation , soil microbiology , bacteria , polymerase chain reaction , chromatography , chemistry , horticulture , gene , biochemistry , bioinformatics , genetics
Aims: To evaluate the suitability of a multiplex PCR‐based assay for sensitive and rapid detection of Escherichia coli O157:H7 in soil and water. Methods and Results: Soil and water samples were spiked with E. coli O157:H7 and subjected to two stages of enrichment prior to multiplex PCR. Detection sensitivities were as high as 1 cfu ml –1 drinking water and 2 cfu g –1 soil. Starvation of E. coli O157:H7 for 35 d prior to addition to soil did not affect the ability of the assay to detect initial cell numbers as low as 10 cfu g –1 soil. Use of an 8‐h primary enrichment enabled detection of as few as 6 cfu g –1 soil, and 10 4 cfu g –1 soil with a 6‐h primary enrichment. When soil was inoculated with 10 5 cfu g –1 , the PCR assay indicated persistence of E. coli O157:H7 during a 35 d incubation. However, when soil was inoculated with lower numbers of pathogen, PCR amplification signals indicated survival to be dependent on cell concentration. Conclusions: A multiplex PCR‐based assay, in combination with an enrichment strategy enabled sensitive and rapid detection of E. coli O157:H7 in soil and water. Significance and Impact of the Study: The ability to sensitively detect E.coli O157:H7 in environmental material within one working day represents a considerable advancement over alternative more time‐consuming methods for detection of this pathogen.