An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef
Abstract The aim of this study is to optimize a rapid and specific method for detection of E. coli O157:H7 (EHEC) from food samples with high background flora using an immunomagnetic separation-real-time polymerase chain reaction assay (IMS-RTiPCR). For this, EHEC cells were recovered from raw milk...
Main Authors: | , |
---|---|
Format: | Article |
Language: | English |
Published: |
Sociedade Brasileira de Ciência e Tecnologia de Alimentos
|
Series: | Food Science and Technology |
Subjects: | |
Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612019005011102&lng=en&tlng=en |
id |
doaj-20452b941c474dd3a244a69fbcd2d5f6 |
---|---|
record_format |
Article |
spelling |
doaj-20452b941c474dd3a244a69fbcd2d5f62020-11-25T02:46:31ZengSociedade Brasileira de Ciência e Tecnologia de AlimentosFood Science and Technology1678-457X010.1590/fst.15818S0101-20612019005011102An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beefBirce MERCANOGLU TABANSait Aykut AYTACAbstract The aim of this study is to optimize a rapid and specific method for detection of E. coli O157:H7 (EHEC) from food samples with high background flora using an immunomagnetic separation-real-time polymerase chain reaction assay (IMS-RTiPCR). For this, EHEC cells were recovered from raw milk and ground meat samples that were artificially contaminated to the final concentrations of 101, 103 and 105 cfu EHEC/mL or g, after a non-selective pre-enrichment at 35 °C for 8 h following either with or without capturing by micro-sized beads. Then, EHEC cells were identified by RTiPCR. The study was also carried out without any enrichment to evaluate the enrichment efficiency of the assay. By comparing the assay with and without pre-enrichment, we showed that even the usage of specific micro-sized beads did not improve EHEC detection in raw milk and ground beef samples and so gave false negative results unless an 8-h pre-enrichment was applied. Besides, the assay can be completed in less than 9 h with a minimum detection limit of 103 cfu EHEC/mL or g in foods and hence was found to be simple routine-based method that exhibits great potential for implementing as a rapid screening of EHEC in microbial tests as compared to existing techniques.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612019005011102&lng=en&tlng=enEscherichia coli 0157:H7immunomagnetic separationmilkraw ground beefreal-time PCR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Birce MERCANOGLU TABAN Sait Aykut AYTAC |
spellingShingle |
Birce MERCANOGLU TABAN Sait Aykut AYTAC An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef Food Science and Technology Escherichia coli 0157:H7 immunomagnetic separation milk raw ground beef real-time PCR |
author_facet |
Birce MERCANOGLU TABAN Sait Aykut AYTAC |
author_sort |
Birce MERCANOGLU TABAN |
title |
An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef |
title_short |
An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef |
title_full |
An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef |
title_fullStr |
An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef |
title_full_unstemmed |
An evaluation of immunomagnetic separation-real-time PCR (IMS-RTiPCR) combined assay for rapid and specific detection of Escherichia coli O157:H7 in raw milk and ground beef |
title_sort |
evaluation of immunomagnetic separation-real-time pcr (ims-rtipcr) combined assay for rapid and specific detection of escherichia coli o157:h7 in raw milk and ground beef |
publisher |
Sociedade Brasileira de Ciência e Tecnologia de Alimentos |
series |
Food Science and Technology |
issn |
1678-457X |
description |
Abstract The aim of this study is to optimize a rapid and specific method for detection of E. coli O157:H7 (EHEC) from food samples with high background flora using an immunomagnetic separation-real-time polymerase chain reaction assay (IMS-RTiPCR). For this, EHEC cells were recovered from raw milk and ground meat samples that were artificially contaminated to the final concentrations of 101, 103 and 105 cfu EHEC/mL or g, after a non-selective pre-enrichment at 35 °C for 8 h following either with or without capturing by micro-sized beads. Then, EHEC cells were identified by RTiPCR. The study was also carried out without any enrichment to evaluate the enrichment efficiency of the assay. By comparing the assay with and without pre-enrichment, we showed that even the usage of specific micro-sized beads did not improve EHEC detection in raw milk and ground beef samples and so gave false negative results unless an 8-h pre-enrichment was applied. Besides, the assay can be completed in less than 9 h with a minimum detection limit of 103 cfu EHEC/mL or g in foods and hence was found to be simple routine-based method that exhibits great potential for implementing as a rapid screening of EHEC in microbial tests as compared to existing techniques. |
topic |
Escherichia coli 0157:H7 immunomagnetic separation milk raw ground beef real-time PCR |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612019005011102&lng=en&tlng=en |
work_keys_str_mv |
AT bircemercanoglutaban anevaluationofimmunomagneticseparationrealtimepcrimsrtipcrcombinedassayforrapidandspecificdetectionofescherichiacolio157h7inrawmilkandgroundbeef AT saitaykutaytac anevaluationofimmunomagneticseparationrealtimepcrimsrtipcrcombinedassayforrapidandspecificdetectionofescherichiacolio157h7inrawmilkandgroundbeef AT bircemercanoglutaban evaluationofimmunomagneticseparationrealtimepcrimsrtipcrcombinedassayforrapidandspecificdetectionofescherichiacolio157h7inrawmilkandgroundbeef AT saitaykutaytac evaluationofimmunomagneticseparationrealtimepcrimsrtipcrcombinedassayforrapidandspecificdetectionofescherichiacolio157h7inrawmilkandgroundbeef |
_version_ |
1724757751985340416 |