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축산가공품의 Listeria monocytogenes 정량적 위해평가에 관한 연구
학
술
Application of Real-time PCR for quantification of Listeria monocytogenes in
발표제목
발
Meat and Milk Processed Products
표
?
Eun Jeong Heo, Hyo-Jin Shin, Young Jo Kim, Hyun Jung Park,
발표자
Hyun Joong Kim, Tae Jun Yoo, Sung Hwan Wee, Jin San Moon
국
내
발표처
한국식품안전성학회
발표시기
2011년 10월
?
수록잡지
(권, 호) 기재
Abstract：
Listeria monocytogenes is a food-born pathogen of concern for the industries,
which has the potential of human listeriosis. It is a ubiquitous microorganism which
is commonly isolated from foods of animal origin like raw milk, meat, meat poultry,
fish and livestock product like ham, sausage, ground meat and milk product and
vegetable and vegetable origin like salad. The purpose of this study was to develop
real-time PCR assays for the quantitative detection of L. monocytogenes organism
in meat and milk processed products. 10g or 10㎖ of ham, sausage, ground meat,
dry sliced meat, processed milk and powdered formula samples and 49 ㎖ of fraser
broth were added in six filtered stomacher bags. For artificial infection, L.
monocytogenes ATCC 19115 was cultured in 37℃ incubator for 24 hr, then 10-fold
5
serially diluted, with final concentration of 10
～10
CFU/㎖. Each dilution was
inoculated in stomacher bags and homogenized in stomacher for 60 sec. All
homogenized samples were transferred to glass bottle and incubated 37℃ incubator
for 3 hr. Then, 1 ㎖ of each homogenized sample and serially diluted L. monocytogenes
was added in eppendorf tube, respectively and used to extract DNA by PrepSeq
rapid spin sample preparation kit (ABI,USA). Realtime PCR for construction of L.
monocytogenes standard curve was performed with designed oligonucleotide primers
and probe pair for the L. monocytogenes inlA gene. When L. monocytogenes were
artificially inoculated in10g samples, the cell concentration of range was
4
approximately 3.5X10
to 3.0X10
CFU/g. The standard curve of serially
diluted
1
5
cells was loglinear for five orders of magnitude from 10
to 10
CFU/㎖ of L.
monocytogenes. When six types of food matrices were artificially contaminated with
4
serially cells of L. monocytogenes, the detection range was from 10
to 3 CFU/g
4
for ham and sausage samples, and from 10
to 10 CFU/g for ground processed
4
2
meat and dry sliced meat samples, and from 10
to 10
CFU/g for processed milk
and powdered formula, respectively. The efficiency of the reaction for ham, sausage,
721
축산가공품의 Listeria monocytogenes 정량적 위해평가에 관한 연구
학
술
Application of Real-time PCR for quantification of Listeria monocytogenes in
발표제목
발
Meat and Milk Processed Products
표
?
Eun Jeong Heo, Hyo-Jin Shin, Young Jo Kim, Hyun Jung Park,
발표자
Hyun Joong Kim, Tae Jun Yoo, Sung Hwan Wee, Jin San Moon
국
내
발표처
한국식품안전성학회
발표시기
2011년 10월
?
수록잡지
(권, 호) 기재
Abstract：
Listeria monocytogenes is a food-born pathogen of concern for the industries,
which has the potential of human listeriosis. It is a ubiquitous microorganism which
is commonly isolated from foods of animal origin like raw milk, meat, meat poultry,
fish and livestock product like ham, sausage, ground meat and milk product and
vegetable and vegetable origin like salad. The purpose of this study was to develop
real-time PCR assays for the quantitative detection of L. monocytogenes organism
in meat and milk processed products. 10g or 10㎖ of ham, sausage, ground meat,
dry sliced meat, processed milk and powdered formula samples and 49 ㎖ of fraser
broth were added in six filtered stomacher bags. For artificial infection, L.
monocytogenes ATCC 19115 was cultured in 37℃ incubator for 24 hr, then 10-fold
5
serially diluted, with final concentration of 10
～10
CFU/㎖. Each dilution was
inoculated in stomacher bags and homogenized in stomacher for 60 sec. All
homogenized samples were transferred to glass bottle and incubated 37℃ incubator
for 3 hr. Then, 1 ㎖ of each homogenized sample and serially diluted L. monocytogenes
was added in eppendorf tube, respectively and used to extract DNA by PrepSeq
rapid spin sample preparation kit (ABI,USA). Realtime PCR for construction of L.
monocytogenes standard curve was performed with designed oligonucleotide primers
and probe pair for the L. monocytogenes inlA gene. When L. monocytogenes were
artificially inoculated in10g samples, the cell concentration of range was
4
approximately 3.5X10
to 3.0X10
CFU/g. The standard curve of serially
diluted
1
5
cells was loglinear for five orders of magnitude from 10
to 10
CFU/㎖ of L.
monocytogenes. When six types of food matrices were artificially contaminated with
4
serially cells of L. monocytogenes, the detection range was from 10
to 3 CFU/g
4
for ham and sausage samples, and from 10
to 10 CFU/g for ground processed
4
2
meat and dry sliced meat samples, and from 10
to 10
CFU/g for processed milk
and powdered formula, respectively. The efficiency of the reaction for ham, sausage,
721
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