8/16/2019 e.coli Pada Bahan Baku Makanan toksik

1/6

See discussions, stats, and author profiles for this publication at: http://www.researchgate.net/publication/6663508

Screening food raw materials for the presenceof the world's most frequent clinical cases of Shiga toxin-encoding Escherichia coli O26,O103, O111, O145 and O157

 ARTICLE  in  INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY · MARCH 2007

Impact Factor: 3.08 · DOI: 10.1016/j.ijfoodmicro.2006.08.014 · Source: PubMed

CITATIONS

70

READS

34

4 AUTHORS, INCLUDING:

Sylvie Perelle

Agence Nationale de Sécurité Sanitaire de l…

53 PUBLICATIONS  1,473 CITATIONS 

SEE PROFILE

Patrick Fach

Agence Nationale de Sécurité Sanitaire de l…

90 PUBLICATIONS  2,723 CITATIONS 

SEE PROFILE

Available from: Patrick Fach

Retrieved on: 09 October 2015

http://www.researchgate.net/profile/Sylvie_Perelle?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_4http://www.researchgate.net/?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_1http://www.researchgate.net/profile/Patrick_Fach?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_7http://www.researchgate.net/institution/Agence_Nationale_de_Securite_Sanitaire_de_lAlimentation_de_lEnvironnement_et_du_Travail?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_6http://www.researchgate.net/profile/Patrick_Fach?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_5http://www.researchgate.net/profile/Patrick_Fach?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_4http://www.researchgate.net/profile/Sylvie_Perelle?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_7http://www.researchgate.net/institution/Agence_Nationale_de_Securite_Sanitaire_de_lAlimentation_de_lEnvironnement_et_du_Travail?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_6http://www.researchgate.net/profile/Sylvie_Perelle?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_5http://www.researchgate.net/profile/Sylvie_Perelle?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_4http://www.researchgate.net/?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_1http://www.researchgate.net/publication/6663508_Screening_food_raw_materials_for_the_presence_of_the_world%27s_most_frequent_clinical_cases_of_Shiga_toxin-encoding_Escherichia_coli_O26_O103_O111_O145_and_O157?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_3http://www.researchgate.net/publication/6663508_Screening_food_raw_materials_for_the_presence_of_the_world%27s_most_frequent_clinical_cases_of_Shiga_toxin-encoding_Escherichia_coli_O26_O103_O111_O145_and_O157?enrichId=rgreq-ed7325ec-5800-49ac-8eab-0cbe20f668eb&enrichSource=Y292ZXJQYWdlOzY2NjM1MDg7QVM6MTAyMDQ1MDQ2NTQyMzQwQDE0MDEzNDA4NDM0NDE%3D&el=1_x_2

8/16/2019 e.coli Pada Bahan Baku Makanan toksik

2/6

Screening food raw materials for the presence of the world's most frequent 

clinical cases of Shiga toxin-encoding  Escherichia coli

O26, O103, O111, O145 and O157

Sylvie Perelle, Françoise Dilasser, Joël Grout, Patrick Fach ⁎

 AFSSA, Agence Française de Sécurité Sanitaire des Aliments (French Agency for Food Safety),

 Laboratoire d'Etudes et de Recherches sur la Qualité des Aliments et sur les Procédés Agroalimentaires (LERQAP),

Unité EBA: Etude moléculaire des contaminants biologiques alimentaires, 23 avenue du Général de Gaulle, 94700 Maisons-Alfort, France

Received 3 November 2005; received in revised form 28 April 2006; accepted 12 August 2006

Abstract

This work aims to provide a strategy for rapidly screening food raw materials of bovine origin for the presence of the most frequent O-

serogroups of Shiga toxin-encoding   Escherichia coli   (STEC) involved in food poisoning outbreaks. The prevalence of highly pathogenic

serogroups of STEC was surveyed in 25 g portions of minced meat and raw milk using PCR-ELISA and multiplex real-time PCR assays. The

 prevalence of STEC in raw milk (n =205) and meat samples (n =300) was 21% and 15%, respectively. Contamination by the main pathogenic

 E. coli O-serogroups representing a major public health concern, including O26, O103, O111, O145, and O157, was potentially around 2.6% in

minced meat and 4.8% in raw milk. The MPN values showed an overall contamination ranging from 1 to 2 MPN cells from highly pathogenic

serogroups/kg. This survey would indicate that the human pathogenic potential of STEC present in these samples probably remains limited. No

conclusion can be drawn at the moment concerning a potential risk for consumers. This rapid screening approach for evaluating the potential

 presence of highly pathogenic serogroups of STEC in food raw materials should help to improve risk assessment of food poisoning outbreaks.© 2006 Elsevier B.V. All rights reserved.

 Keywords:  STEC; O-serogroup; Real-time PCR; Food testing

1. Introduction

Since the early 1980s, Shiga toxin-producing   Escherichia

coli   (STEC) have emerged as a major cause of food-borne

infections (Karmali et al., 1983; Riley et al., 1983). STEC can

cause diarrhea and hemorrhagic colitis (HC) and may lead to

haemolytic uraemic syndrome (HUS). In Europe, HUS is now believed to be the commonest cause of acute renal failure in

children, and may be fatal in up to 10% of cases (Padhye and

Doyle, 1992). Infections may result from direct contact with

animals or their faeces (Locking et al., 2001) but most human

infections originate from the consumption of contaminated

water or raw or undercooked food. A range of different 

foodstuffs including vegetables, salad bar items, fruit, salami,

and fresh cheese made of raw goats' and cows' milk have been

epidemiologically implicated in serotype O157 outbreaks

(Besser et al., 1993; CDC, 1995; Del Rosario and Beuchat,

1995; Deschenes et al., 1996). Nevertheless, raw meat, in

 particular beef, has been considered as the principal source of 

STEC (Karmali, 1989), and ruminants are still considered the

major STEC reservoir. While numerous outbreaks have been

attributed to STEC strains of serotype O157:H7, it has beenestablished that STEC strains causing gastrointestinal diseases

in humans may well belong to more than 100 serotypes

(Karmali, 1989; Russmann et al., 1995; Paton and Paton, 1998;

 Nataro and Kaper, 1998). However, the value of screening for 

the presence of the   stx  gene sequences remains controversial,

since not all STEC prove to be clinically significant in humans.

A better understanding of the scientific basis for the difference

in virulence between serotypes would help to identify specific

targets characterizing strains that pose a significant risk in

human diseases. However, although certain differences in viru-

lence between groups of STEC strains have been related to the

International Journal of Food Microbiology 113 (2007) 284 –288

www.elsevier.com/locate/ijfoodmicro

⁎  Corresponding author. Tel.: +33 1 4977 2813; fax: +33 1 4368 9762.

 E-mail address: p.fach@afssa.fr  (P. Fach).

0168-1605/$ - see front matter © 2006 Elsevier B.V. All rights reserved.doi:10.1016/j.ijfoodmicro.2006.08.014

http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-mailto:p.fach@afssa.frhttp://dx.doi.org/10.1016/j.ijfoodmicro.2006.08.014http://dx.doi.org/10.1016/j.ijfoodmicro.2006.08.014mailto:p.fach@afssa.frhttp://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-

8/16/2019 e.coli Pada Bahan Baku Makanan toksik

3/6

 presence of specific pathogenicity islands (PAIs), e.g. the locus

of enterocyte effacement (LEE), the scientific bases for as-

sessing the potential virulence of STEC have not yet been

elucidated (Karmali et al., 2003). Several reports have

mentioned O26, O103, O111, O145 and O157:H7 as the most 

frequent O-serogroups involved in food poisoning outbreaks,

leading STEC strains from these O-serogroups to be consideredas the world's main pathogenic strains with implications for 

 public health. Despite several real-time PCR O-serotyping

assays are available to rapidly determine certain STEC

serogroups (Sharma, 2002; Perelle et al., 2004, 2005), surveys

in foods have rarely focused on these five major serogroups. It is

extremely important to evaluate natural contamination of raw

milk and beef meat by STEC O26, STEC O103, STEC O111,

STEC O145 and STEC O157:H7 for efficient risk assessment of 

food poisoning outbreaks, but there are still few prevalence data

available.

The aim of this study was to evaluate the prevalence of these

STEC O-serogroups in minced meat and raw milk samplescollected in France. The study method is based on enrichment of 

samples in a nutrient broth before positive STEC screening

using a recently developed duplex 5′-nuclease PCR assay

(Perelle et al., 2004) targeting the   stx   genes, followed by a

newly developed multiplex 5′-nuclease PCR test specific for 

 E. coli   O26,   E. coli   O103,   E. coli   O111,   E. coli   O145 and

 E. coli O157. The advantages and disadvantages of using such a

strategy for foods analysis are discussed.

2. Materials and methods

2.1. Food samples

More than five hundred samples of bovine origin were

collected in France from different geographical areas across

the country during a period of 12 months. They arrived at the

laboratory by rapid cold road transport and were processed on

arrival. The samples of food raw materials were raw milk 

(n =205) collected in several dairies and minced meat (n =300)

collected in different abattoirs. Beef meat was cut using sterile

instruments, placed in individual plastic bags, homogenized

in a stomacher blender for 120 s, and then stored at  −20 °C

until use. Raw milk samples were stored at 4 °C. The sample

quantities prepared were approximately 25 g for each sam-

 ple. The samples were stored not longer than 48 h beforeinvestigation.

2.2. Enrichment procedure and DNA extraction

Samples of 25 g of raw material were diluted tenfold (wt/vol)

in 225 ml of modified EC medium containing 20 mg l− 1 of 

novobiocin (mEC+n broth), and incubated at 37 °C overnight.

After incubation, a 1 ml aliquot of the enrichment broth was

collected for DNA extraction. This aliquot was centrifuged for 

5 min at 9000  g , and the supernatant was discarded. The cell

 pellets were then DNA-extracted using InstaGene Matrix™

(Bio-Rad Laboratories, Marnes-La-Coquette, France) and the

DNA was stored at  −20 °C until PCR testing.

2.3. Food PCR detection

To detect STEC in food samples, we used the PCR-ELISA

described previously (Fach et al., 2001). Briefly, four 

degenerated oligonucleotides targeting the most well-con-

served sequences in the  stx  genes were designed for screening

 stx1   and   stx2   gene sequences in food products (Fach et al.,2001). Two oligonucleotides were designed as primers to

amplify approximately 500 bp fragments from  stx  genes, while

the two others were used as capture and detection probes in a

sandwich hybridization assay. The capture probe was labelled

with biotin and bound to streptavidin-coated microtiter plates,

while the detection probe was labelled with digoxigenin. After 

alkaline denaturation of the PCR products, hybridization oc-

curred with the two probes in the streptavidin-coated microtiter 

 plates. In a similar way to ELISA technology, hybrids were

detected using a peroxidase anti-digoxigenin conjugate, and

the final enzymatic reaction of the test gave a colorimetric

signal measured with a microtiter plate reader. To identify possible PCR inhibitors derived from the food samples, an

internal amplification control (IAC) was included which co-

amplified with stx  target genes. PCR products derived from the

IAC were detected with a specific set of capture and detection

 probes (Fach et al., 2001). The PCR-ELISA positive samples

were further tested in a newly developed multiplex real-time

PCR assay screening for   E. coli   O26,   E. coli   O103,   E. coli

O111, E. coli O145 and E. coli O157. This multiplex PCR test 

is based on primers and probes targeting the O-antigen gene

clusters of  E. coli O26, E. coli O111 and E. coli O157, the eae

gene of  E. coli O103, and the O-island 29 of  E. coli  O145, as

described in a previous study (Perelle et al., 2004). Multiplex

amplifications were performed using 20  μl of reaction mixturecontaining 1× concentration of a LightCycler-Faststart DNA

master hybridization probes mix (Roche Diagnostics, Meylan,

France), 5 mM MgCl2, 500 nM of each primer, 200 nM of each

FAM-TAMRA labelled probe, and 2  μl of the template DNA.

Positive controls using appropriate purified plasmid DNA

(Perelle et al., 2004) and two negative controls containing

all the reagents except the DNA template were included

with each amplification set. Cycling was carried out in the

LightCycler instrument (Roche Diagnostics). The amplifica-

tion started with an initial denaturing step at 95 °C for 10 min,

followed by 40 cycles at 95 °C for 10 s with a temperature

transition rate of 20 °C/s, and 60 °C for 30 s with a temperaturetransition rate of 10 °C/s, and then a cooling step at 40 °C for 

30 s. The generation of fluorescence for each sample was

monitored at the end of the elongation steps in the F1/F2

channel (530 nm). A fluorescent signal 10-fold higher than the

standard deviation of the mean baseline emission indicated a

 positive detection. Ct was defined as the PCR cycle at which

the fluorescent intensity exceeds the threshold. Finally,

samples tested positive by the multiplex real-time PCR assay

were further tested in simplex real-time PCR assays to identify

the exact pathogenic   E. coli   O-serogroups concerned, as

well as in an  stx-typing 5′-nuclease PCR assay (Perelle et al.,

2004). The method proposed in our study is summarized

in Fig. 1.

285S. Perelle et al. / International Journal of Food Microbiology 113 (2007) 284 – 288

http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-

8/16/2019 e.coli Pada Bahan Baku Makanan toksik

4/6

2.4. Determination of MPN counts

The survey data were converted to MPN counts as

 previously proposed (Halvorson and Ziegler, 1933). MPNs

were calculated using the following formula: MPN=Ln(n / q),

where  n   is the number of samples tested and  q  the number of 

negative samples.

3. Results and discussion

A total of 505 food raw materials of bovine origin were

screened by PCR for the presence of the most frequent O-

serogroups of STEC involved in food poisoning outbreaks.

Screening of STEC was performed by PCR-ELISA with an IAC

to monitor false negative results due to sample PCR inhibitors

(Fach et al., 2001). STEC-positive samples were further tested

in the newly developed multiplex real-time PCR assay for 

 E. coli   O26,   E. coli   O103,   E. coli   O111,   E. coli   O145 and

 E. coli  O157. Finally, samples tested positive by the multiplex

real-time PCR assay were further tested in simplex real-time

PCR assays to identify the exact pathogenic   E. coli   O-

serogroups concerned, as well as in an   stx-typing 5′-nucleasePCR assay (Perelle et al., 2004).

The prevalence of STEC-positive samples as determined by

PCR-ELISA was 17.4% (Table 1). Forty-five (15%) of the 300

samples of beef meat and 43 (21%) of the 205 samples of raw

milk were positive for STEC in PCR-ELISA. Seventy-four 

(84%) of the 88 positive samples were confirmed positive by

 stx-typing with 5′-nuclease PCR assay. This difference between

the PCR-ELISA and 5′-nuclease PCR may be due either to the

Table 1

Evaluation of the prevalence of STEC O26, O103, O111, O145 and O157 in food raw materials of bovine origin

Sample type Number 

tested

 Number of positive samples Most 

 probable

number/ 

kg a 

PCR-ELISA Typing of the stx  genes by 5′-nuclease PCR Specific O-serogroup identification of STEC-positive samples

 Number 

(%)

 Number 

(%)

(Number of samples)

Minced meat 

(beef)

300 45 (15%) 2  stx1 (4.6%), 28  stx2  (65%),

and 13  stx1+ stx2  (30.2%)

O145 (4), O157 (1), O145+O157 (1), O145+O103 (1),

and O157+O103 (1)

1

(Total=8)

Raw milk 205 43 (21%) 12  stx1  (38.7%), 10  stx2  (32.2%),

and 9  stx1+ stx2 (29%)

O145 (4), O103 (1), O145+O157 (3), O145+O103 (1),

and O145+O26 (1)

2

(Total=10)

Total 505 88 (17.4%) 14  stx1  (18.9%), 38  stx2  (51.3%),

and 22  stx1+ stx2  (29.7%)

O145 (8), O157 (1), O103 (1), O145+O157 (4),

O145+O103 (2), O145+O26 (1), and O157+O103 (1)

1–2

(Total = 74) (Total = 18)

a  According to Halvorson and Ziegler (1933).

Fig. 1. Flow diagram illustrating the preparation of food samples and the different screening steps for evaluating the presence of STEC serogroups of major public

health concern.

286   S. Perelle et al. / International Journal of Food Microbiology 113 (2007) 284 – 288

http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-

8/16/2019 e.coli Pada Bahan Baku Makanan toksik

5/6

different quantities of DNA tested in the two PCR tests (10 μl in

PCR-ELISA against 2  μl in real-time PCR), to the presence of 

PCR inhibitors in the samples which may have affected each

PCR system differently, or to possible STEC variants that 

cannot be equally detected with the two PCR systems, which

would be in agreement with the increasing number of variants

described in the literature over the last few years (Gyles et al.,

1988; Weinstein et al., 1988; Gannon et al., 1990; Ito et al.,

1990; Hii et al., 1991; Lin et al., 1993; Kim et al., 1997; Paton

et al., 1992, 1993a,b, 1995; Pierard et al., 1998). These variants

have usually been described as Stx2 strains of STEC, while

variants derived from Stx1 strains have rarely been reported

(Paton et al., 1993a, 1995). 5′-nuclease PCR typing of the  stx

genes demonstrated that 14 (19%) samples carried  stx1  genes,

38 (51.3%) carried stx2 genes, and 22 (29.7%) carried both  stx1and stx2 genes (Table 1).

Multiplex real-time PCR detection of   E. coli  O26,  E. coli

O103, E. coli  O111,  E. coli  O145 and  E. coli  O157 confirmed

only 18 positives out of the 74 positives obtained by the 5′-

nuclease PCR targeting the   stx   genes. This suggests that the

overall contamination by the main pathogenic   E. coli   O-

serogroups of major public health concern was around 3.5% in

these food samples, broken down as 2.6% in minced meat and

4.8% in raw milk. The MPN values would indicate an overall

 potential contamination ranging from 1 to 2 MPN STEC cells of highly pathogenic serogroup/kg, meaning that contamination of 

 beef meat and raw milk by highly pathogenic serogroups of 

STEC is very low, and that the risk of consumer infection by

human pathogenic strains of STEC present in these samples is

 probably very minor.

The 18 samples tested positive in multiplex real-time PCR 

were identified with high precision by simplex real-time PCR 

(Table 1). Eight samples were positive with   E. coli   O145, 1

sample was positive with  E. coli  O103, 1 sample was positive

with   E. coli  O157, 4 samples were positive with both   E. coli

O145 and   E. coli   O157, 2 samples were positive with both

 E. coli  O103 and E. coli O145, 1 sample was positive with both

 E. coli  O103 and E. coli O157, and 1 sample was positive with

 both   E. coli   O26 and   E. coli   O145. None of the samples

analyzed tested positive with   E. coli   O111. The overall

contamination of beef meat and raw milk by   E. coli   O157

was 1%, which is in strong agreement with the reported

worldwide E. coli  O157 incidence in these food products. The

 prevalence of  E. coli  O26 and  E. coli  O103 was very low at 1

and 3 positive samples, respectively, which is in the lower range

of worldwide reported incidence for these two serotypes. The

 prevalence of   E. coli  O145 (positive in 3% of samples) was

unexpectedly higher than the prevalence of other serogroups.

This study demonstrates that different O-serogroups of 

 E. coli   of major public health concern are able to enter the

 processing chain in the meat and dairy industries. However,there is increasing evidence in the literature that many of these

STEC O-serogroups may not produce Shiga-toxin, nor will

they posses the  stx genes. Fig. 2 illustrates that both toxigenic

( stx-positive) and non-toxigenic ( stx-negative) strains coex-

isted within each O-serogroup. Therefore, when both  stx  and

O-serogroup gene sequences were detected in certain foods

(2.6% of minced meat and 4.8% of raw milk), there was no

evidence that these signals were displayed by a pathogenic  E.

coli strain. As mentioned in Fig. 1, isolation of clones from the

food mixture should be performed to confirm or notthe positive

 presumptive result. But, this isolation step is still problematic

and time-consuming using the current commercially availablemedia. Whatever the difficulties to isolate STEC from foods,

the idea of combining detection of  stx and genes encoding O-

groups is a good approach along our way to optimise methods

for detection of pathogenic STEC. The rapid (within 36 h)

detection and typing of the  stx  and O-serogroups of STEC in

food raw materials should be helpful in forming a response to

 E. coli  food poisoning.

Acknowledgment

This work was partially funded by  Ministère de l'Agriculture

et de la Pêche, Direction Générale de l'Alimentation   (French

Ministry for Agriculture and Fisheries), Paris, France.

Fig. 2. Diagram showing the coexistence of both toxigenic ( stx-positive) and non-toxigenic ( stx-negative) strains within each O-serogroup tested.

287S. Perelle et al. / International Journal of Food Microbiology 113 (2007) 284 – 288

http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-http://-/?-

8/16/2019 e.coli Pada Bahan Baku Makanan toksik

6/6

References

Besser, R.E., Lett, S.M., Weber, J.T., Doyle, M.P., Barett, T.J., Wells, J.G.,

Griffin, P.M., 1993. An outbreak of diarrhea and haemolytic uremic

syndrome from   Escherichia coli  O157:H7 in fresh-pressed apple cider.

Journal of the American Medical Association 269, 2217–2220.

Centers for Disease Control, 1995. Escherichia coli O157:H7 outbreak linked to

commercially distributed dry-cured salami  —  Washington and California,1994. Morbidity and Mortality Weekly Report 44, 157–160.

Del Rosario, B.A., Beuchat, L.R., 1995. Survival and growth of entero-

hemorrhagic   Escherichia coli   O157:H7 in cantaloupe and watermelon.

Journal of Food Protection 58, 105–107.

Deschenes, G., Casenave, C., Grimont, F., Desenclos, J.C., Benoit, S., Collin,

M., Baron, S., Mariani, P., Grimont, P.A., Nivet, H., 1996. Cluster of cases of 

haemolytic uraemic syndrome due to unpasteurised cheese. Pediatric

 Nephrology 10, 203–205.

Fach, P., Perelle, S., Dilasser, F., Grout, J., 2001. Comparison between a PCR-

ELISA test and the Vero cell assay for detecting Shiga toxin-producing

 Escherichia coli   in dairy products and characterization of virulence traits

of the isolated strains. Journal of Applied Microbiology 90, 809–818.

Gannon, V.P.J., Teerling, C., Masri, S.A., Gyles, C.L., 1990. Molecular cloning

and nucleotide sequence of another variant of the  Escherichia coli   Shiga-

like toxin II family. Journal of General Microbiology 136, 1125–1135.

Gyles, C.L., De Grandis, S.A., MacKenzie, C., Brunton, J.L., 1988. Cloning and

nucleotide sequence analysis of the genes determining verocytotoxin

 production in a porcine edema disease isolate of  Escherichia coli. Microbial

Pathogenesis 5, 419–426.

Halvorson, H.O., Ziegler, N.R., 1933. Application of statistics to problems in

 bacteriology. I. A means of determining bacterial population by the dilution

method. Journal of Bacteriology 25, 101–121.

Hii, J.H., Gyles, C., Morooka, T., Karmali, M.A., Clarke, R., De Grandis, S.,

Brunton, J.L., 1991. Development of verotoxin 2-and verotoxin 2 variant 

(VT2v)-specific oligonucleotide probes on the basis of the nucleotide

sequence of the B cistron of VT2v from  Escherichia coli E32511 and B2F1.

Journal of Clinical Microbiology 29, 2704–2709.

Ito, H., Terai, A., Kurazono, H., Takeda, Y., Nishibuchi, M., 1990. Cloning and

nucleotide sequencing of Vero toxin 2 variant genes from  Escherichia coli

O91:H21 isolated from a patient with the hemolytic uremic syndrome.

Microbial Pathogenesis 8, 47–60 (Published erratum appears in Microbial

Pathogenesis 8, 449).

Karmali, M.A., 1989. Infection by verocytotoxin-producing  Escherichia coli.

Clinical Microbiology Reviews 2, 15–38.

Karmali, M.A., Steele, B.T., Petric, M., Lim, C., 1983. Sporadic cases of 

haemolytic-uraemic syndrome associated with faecal cytotoxin and

cytotoxin-producing  Escherichia coli   in stools. Lancet 1, 619–620.

Karmali, M.A., Mascarenhas, M., Shen, S., Ziebell, K., Johnson, S., Reid-Smith,

R., Isaac-Renton, J., Clark, C., Rahn, K., Kaper, J.B., 2003. Association of 

genomic O island 122 of  Escherichia coli   EDL 933 with verocytotoxin-

 producing Escherichia coli  seropathotypes that are linked to epidemic and/ 

or serious disease. Journal of Clinical Microbiology 41, 4930–4940.

Kim, S.H., Cha, I.H., Kim, K.S., Kim, Y.H., Lee, Y.C., 1997. Cloning and

sequenceanalysis of another Shiga-like toxin IIe variant gene (slt-IIera) from

an   Escherichia coli  R107 strain isolated from rabbit. Microbiology and

Immunology 41, 805–808.

Lin, Z., Yamasaki, S., Kurazono, H., Ohmura, M., Karasawa, T., Inoue, T.,

Sakamoto, S., Suganami, T., Takeoka, T., Taniguchi, Y., 1993. Cloning and

sequencing of two new Verotoxin 2 variant genes of   Escherichia coli

isolated from cases of human and bovine diarrhea. Microbiology and

Immunology 37, 451–459.

Locking, M.E., O'Brien, S.J., Reilly, W.J., Wright, E.M., Campbell, D.M., Coia,

J.E., Browning, L.M., Ramsay, C.N., 2001. Risk factors for sporadic cases

of  Escherichia coli  O157 infection: the importance of contact with animal

excreta. Epidemiology and Infection 127, 215–220.

 Nataro, J.P., Kaper, J.B., 1998. Diarrheagenic   Escherichia coli. ClinicalMicrobiology Reviews 11, 142–201 (published erratum appears in Clinical

Microbiology Reviews 11, 403).

Padhye, N.V., Doyle, M.P., 1992.   Escherichia coli  O157:H7 epidemiology,

 pathogenesis and methods for detection in food. Journal of Food Protection

55, 555–565.

Paton, J.C., Paton, A.W., 1998. Pathogenesis and diagnosis of Shiga toxin-

 producing Escherichia coli   infections. Clinical Microbiology Reviews 11,

450–479.

Paton, A.W., Paton, J.C., Heuzenroeder, M.W., Goldwater, P.N., Manning, P.A.,

1992. Cloning and nucleotide sequence of a variant Shiga-like toxin II gene

from Escherichia coli OX3:H21 isolated from a case of sudden infant death

syndrome. Microbial Pathogenesis 13, 225–236.

Paton, A.W., Paton, J.C., Goldwater, P.N., Heuzenroeder, M.W., Manning, P.A.,

1993a. Sequence of a variant Shiga-like toxin type-I operon of  Escherichia

coli  O111:H−

. Gene 129, 87–92.Paton, A.W., Paton, J.C., Manning, P.A., 1993b. Polymerase chain reaction

amplification, cloning and sequencing of variant  Escherichia coli Shiga-like

toxin type II operons. Microbial Pathogenesis 15, 77–82.

Paton, A.W., Beutin, L., Paton, J.C., 1995. Heterogeneity of the amino-acid

sequences of  Escherichia coli   Shiga-like toxin type-I operons. Gene 153,

71–74.

Perelle, S., Dilasser, F., Grout, J., Fach, P., 2004. Detection by 5′-nuclease PCR 

of Shiga-toxin producing  Escherichia coli   O26, O55, O91, O103, O111,

O113, O145 and O157:H7, associated with the world's most frequent 

clinical cases. Molecular and Cellular Probes 18, 185–192.

Perelle, S., Dilasser, F., Grout, J., Fach, P., 2005. Detection of   Escherichia coli

serogroup O103 by real-time Polymerase Chain Reaction. Journal of 

Applied. Microbiology 98, 1162–1168.

Pierard, D., Muyldermans, G., Moriau, L., Stevens, D., Lauwers, S., 1998.

Identification of new verocytotoxin type 2 variant B-subunit genes in humanand animal  Escherichia coli   isolates. Journal of Clinical Microbiology 36,

3317–3322.

Riley, L.W., Remis, R.S., Helgerson, S.D., McGee, H.B., Wells, J.G., Davis, B.R.,

Hebert, R.J., Olcott, E.S., Johnson, L.M., Hargrett, N.T., Blake, P.A., Cohen,

M.L., 1983. Hemorrhagic colitis associated with a rare   Escherichia coli

serotype. New England Journal of Medecine 308, 681–685.

Russmann, H., Kothe, E., Schmidt, H., Franke, S., Harmsen, D., Caprioli, A.,

Karch, H., 1995. Genotyping of Shiga-like toxin genes in non-O157

 Escherichia coli   strains associated with haemolytic uraemic syndrome.

Journal of Medical Microbiology 42, 404–410.

Sharma, V.K., 2002. Detection and quantitation of enterohemorrhagic  Escher-

ichia coli   O157, O111, and O26 in beef and bovine feces by real-time

 polymerase chain reaction. Journal of Food Protection 65, 1371–1380.

Weinstein, D.L., Jackson, M.P., Samuel, J.E., Holmes, R.K., O'Brien, A.D.,

1988. Cloning and sequencing of a Shiga-like toxin type II variant from Escherichia coli   strain responsible for edema disease of swine. Journal of 

Bacteriology 170, 4223–4230.

288   S. Perelle et al. / International Journal of Food Microbiology 113 (2007) 284 – 288