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과 제 명 축산가공품의 Listeria monocytogenes 정량적 위해평가에 관한 연구
학
술
Comparison of four DNA extraction methods in the real time PCR assay for
발표제목
발
detection of Listeria monocytogenes from milk products
표
발 표 자 허은정 외 5명
게
재
발 표 처 한국식품위생학회
발표시기
2012년 10월
?
국
내
Abstract :
?
Listeria monocytogenes is important foodborne pathogen, commonly isolated from foods
of animal origin like raw milk, meat, poultry and fish and vegetable and vegetable
origin like salad. Real time PCR assay for rapid detection on pathogen in food have
become choice due to its high sensitivity and specificity. Therefore, the efficiency of
DNA extraction from various food matrices is important to detect and quantify this
organism, especially when this organism is contaminated with low concentration. We
compared the efficiency by evaluating conventional DNA extraction(boiling) method and
three commercially available kits(Qiagen DNase mini kit, ABI rapid spin sample
purification kit and Genall cell SV mini kit) for DNA extraction of L. monocytogenes
cultured solution in the real time PCR assay. L. monocytogenes cultured for 24 h was
5
serially diluted with range from 10
to 10
CFU/㎖ and spiked in milk and milk
products(cheese and milk formula) with cultivation for 4 h at 37 ℃. Spiked and
cultured samples were extracted with each kit and evaluated by detection limit and
standard curve using a TaqMan Listeria monocytogenes detection kit(ABI). The lowest
detection limit was 10
CFU/㎖ in each food matrices, which was obtained with ABI
1
3
and Geneall kit, whereas Qiagen kit and boiling method had limits of 10
- 10
CFU/
㎖. When standard curves compared, Geneall kit was more efficient in three types of
2
food matrices of milk, cheese and infant milk formula with R
of > 0.98 and efficiency
2
of 105-111%, while ABI kit showed R
of 0.85 - 0.98 and efficiency of 119 - 200%.
This result suggested that ABI and Geneall kit could provide the most sensitive
methods in the real time quantification PCR assay for detecting of L. monocytogenes
from milk and milk products.
909
학
술
Comparison of four DNA extraction methods in the real time PCR assay for
발표제목
발
detection of Listeria monocytogenes from milk products
표
발 표 자 허은정 외 5명
게
재
발 표 처 한국식품위생학회
발표시기
2012년 10월
?
국
내
Abstract :
?
Listeria monocytogenes is important foodborne pathogen, commonly isolated from foods
of animal origin like raw milk, meat, poultry and fish and vegetable and vegetable
origin like salad. Real time PCR assay for rapid detection on pathogen in food have
become choice due to its high sensitivity and specificity. Therefore, the efficiency of
DNA extraction from various food matrices is important to detect and quantify this
organism, especially when this organism is contaminated with low concentration. We
compared the efficiency by evaluating conventional DNA extraction(boiling) method and
three commercially available kits(Qiagen DNase mini kit, ABI rapid spin sample
purification kit and Genall cell SV mini kit) for DNA extraction of L. monocytogenes
cultured solution in the real time PCR assay. L. monocytogenes cultured for 24 h was
5
serially diluted with range from 10
to 10
CFU/㎖ and spiked in milk and milk
products(cheese and milk formula) with cultivation for 4 h at 37 ℃. Spiked and
cultured samples were extracted with each kit and evaluated by detection limit and
standard curve using a TaqMan Listeria monocytogenes detection kit(ABI). The lowest
detection limit was 10
CFU/㎖ in each food matrices, which was obtained with ABI
1
3
and Geneall kit, whereas Qiagen kit and boiling method had limits of 10
- 10
CFU/
㎖. When standard curves compared, Geneall kit was more efficient in three types of
2
food matrices of milk, cheese and infant milk formula with R
of > 0.98 and efficiency
2
of 105-111%, while ABI kit showed R
of 0.85 - 0.98 and efficiency of 119 - 200%.
This result suggested that ABI and Geneall kit could provide the most sensitive
methods in the real time quantification PCR assay for detecting of L. monocytogenes
from milk and milk products.
909
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