establishing quantitative evaluation method of contact...
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加藤研究室・大岡研究室・菊本研究室Kato Lab., Ooka Lab., and Kikumoto Lab.
Establishing quantitative evaluation method of contact infection risk using qPCR method
Background /Purpose
1/2
= +
Real-time PCR Thermal cycler MonitoringFluorescent
substance
+
Background
■ Increase of cities’ population and
Processing of building air in recent years
∴Elevation of infection risk→Need to control infection
■ The mechanism of contact infection is complicated (※1)
therefore, the evaluation method is not established yet
■ Development of gene analysis technology and
PCR(※2)method
・It is classified as three major infection routes among several infection routes.
(Air infection, droplet infection, contact infection)
・Frequent occurrence as typified by norovirus
・It is impossible to predict by CFD unlike air infection and droplet infection
・A method developed by Carrie-Maris in 1983.
・It makes analysis faster , accurate and easier. Among them, this
study used real-time PCR method is used in this study.
This is a method of monitoring the production process of a PCR
product using a fluorescent substance and measuring
(quantifying and detecting) the target DNA .(Below figure)
・Adenosine TriPhosphate (: ATP) method has been used as a
commonly used method until now, but this method is very difficult
to measure because it targets cells contained in all organisms.
Purpose
■Establishing quantitative evaluation method of contact infection
risk using qPCR method
・Establish analysis method
・Establish experimental method
■Creation of highly accurate infection simulator by combination
of behavior simulation and simulator (CFD) of other infection
route
※1 Contact infection ※2 PCR(Polymerase Chain Reaction) method
資料 :地理院地図
参考 : http://kansenyobou.net/
➢Infection
routeCharacteristic
Main causes
Microorganisms
Contact
infection
· Frequent propagation path that propagates
through fingers, foods and appliancesNorovirus
Droplet
infection
Coughing, sneezing, conversation, etc.,
propagate by droplet particles
It falls to the floor within 1 m.
It never keeps floating in the air
Influenza virus
Legionella spp.
Airborn
infection
Coughing, sneezing, etc. will propagate as
droplet nuclei.
Floating in the air, scattering by airflow
M. tuberculosis
加藤研究室・大岡研究室・菊本研究室Kato Lab., Ooka Lab., and Kikumoto Lab.
Establishing quantitative evaluation method of contact infection risk using qPCR method
Procedure / Result
2/2
Procedure BackgroundResult
Ap
ply
ing
Dry
ing
Sam
plin
g
·Tracer
・Drying time
・Sampling method
Decora
Float glass
Frosted glass
Lactic acid bacteria(Lactobacillus delbrueckii subsp. Bulgaricus OLL1073R-1)
Dilution rate× 10
× 100
× 1000
1’00”
2’30”
4’00”
5’30”
7’30”
· Experimental location: Kato Laboratory Lab., The Univ. of Tokyo
Wipe off with uniform load three times in each direction
①
④
③
Store at -20 °C and analyze by qPCR method
Experimental flow and conditions in each step
Applying
amount
50μl
100μl
0
2
4
6
8
10
12
14
16
1(n=4)
2.5(n=3)
4(n=3)
5.5(n=5.5)
7.7(n=7.5)
Co
llect
ion
rat
e (%
)
Drying time(minuites)
Average
0
1
2
3
4
5
6
7
8
50(n=3) 100(n=4)
Co
llect
ion
rat
e (%
)
Applying amount(μl)
Average
0
0.5
1
1.5
2
2.5
3
3.5
10(n=4) 100(n=4) 1000(n=4)
Co
llect
ion
rat
e (%
)
Dilution rate
Average
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
Float glass(n=4)
Frosted glass(n=4)
Decora(n=3)
Co
llect
ion
rat
e (%
)
Material name
Average
■Although the deviation was large in all cases, a large correlation with the collection
rate was not observed except for the drying time (④)
■As the collection rate has become very low as a whole, it will be a future task
・Material type
②
Examined the sampling rate when changing the conditions of ①-④
①Collection rate depend on dilution rate ②Collection rate depend on Applying amount
③Collection rate depend on material type ④Collection rate depend on drying time
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