Loopamp Listeria monocytogenes Detection Kit ... - Mast Diagnostica

For the purpose of foods
and environment inspection
Loopamp ® Listeria monocytogenes Detection Kit
•Listeria monocytogenes in foods can be detected in only 2 days.
Listeria monocytogenes can be detected in a short time. Including pre-culture, detection of Listeria
monocytogenes takes only 2 days.
•Rapid and specific detection of Listeria monocytogenes in foods can be achieved.
The detection can be done in a short time due to the high amplification efficiency of LAMP method. By
using 4 primers to recognize 6 distinct regions, high specificity can be achieved.
•From amplification reaction to detection, the whole process is done in 1 step with
Loopamp Realtime Turbidimeters.
From gene amplification to detection, the whole process is done in 1 step, in a closed system (in 1
single tube). Amplification of the target gene can be judged by detecting the white precipitates of
magnesium pyrophosphate, an amplification by-product.
Manufacturer
1-33-8, HONGO, BUNKYO-KU, TOKYO 113-8408 JAPAN

The Assay Principle
Amplification of nucleic acids is conducted by the LAMP method using Listeria monocytogenes invasion gene (iap)
sequence as the primers to detect Listeria monocytogenes. From whether the amplification occurred or not, the
existence of Listeria monocytogenes can be determined. Gene amplification is detected by using Loopamp Realtime
Turbidimeters to monitor the turbidity of white precipitates of magnesium pyrophosphate, which is a by-product of
amplification reaction.
Operation Procedure
Food 25g + 225mL of Half Fraser media, UVM media or EB media
Incubate at 30ºC for 24 hours.
Pipette 50µL of enrichment culture into the tube.
Centrifuge at 2,000Xg for 5 minutes and remove the supernatant.
Add 80µL of EX F. Alkaline heat treatment at 95ºC for 5 minutes.
Preparing the Master Mix (for 1 test)
Reaction Mix. Lis 20µL
Bst DNA Polymerase 1µL
Total 21µL
Add 10µL of 1M Tris-HCl and mix.
Centrifuge at 2,000Xg for 30 seconds.
Use the supernatant as sample.
Dispense 20µL of Master Mix into
each reaction tube.
Add 5µL of Sample Solution or controls into the tubes.
Set the reaction tubes into the reaction blocks of Loopamp Realtime Turbidimeters and start reaction.
LAMP reaction 65ºC, 60 minutes
Enzyme inactivation (at 80ºC for 2 minutes, done by Loopamp Realtime Turbidimeters automatically)
Detection and Judgment
Contents of the kit (for 48 tests)
*When using this kit, use the
specific reaction tubes.
(1)Extraction Solution for Foods (EX F) 1.8mL X 3 tubes
(2)1M Tris-HCl: pH7.0 (Tris) 1.0mL X 1 tube
(3)Reaction Mix. Lis (RM Lis) 1.0mL X 1 tube
(4)Bst DNA Polymerase
(Bst DNA Polymerase) 60µL X 1 tube
(5)Positive Control Lis (PC Lis) 0.1mL X 1 tube
The notation on each reagent tube is shown in ( ).
Loopamp ® Reaction Tube (separately sold)
Unit, Storage, Expiration, Code No.
Product Name
Unit
Storage
Expiration
Loopamp ® Listeria monocytogenes
Detection Kit
Loopamp ® Reaction Tube
48 tests
8 x 12 sets
-20ºC
Room Temperature
1 year
-
* For purchasing and inquiries, contact the following e-mail: international@eiken.co.jp
Code No.
LMP701
LMP903
Manufacturer
1-33-8, HONGO, BUNKYO-KU, TOKYO 113-8408 JAPAN
Dec, 2005