Cica Geneus Pseudo POT KIT
TMis a kind of representative indigenous bacterium duringenvironment. In the hospital-acquired infections, the Multiple-Drug-Resistant（MDRP）which got tolerance for a main antibiotic widely become
the problem.
The PCR-based ORF Typing（POT）*1 developed by Dr.Suzuki of Aichi Prefectural Institute of Public Health and Dr.Iinuma of Kanazawa Medical University is a method toamplify plural speciﬁc genes by Multiplex PCR at the same time, and then to analyze
agarose gel electrophoresis banding patterns.
References
Heisei 23, Health and labour sciences research grants (the H1N1 ﬂu virus etc. emerging and reemerging infectious diseases research project) [Study fornew drug resistant bacteria] Generalization, allotment working papers study, representative Dr.Arakawa.

Characteristics●Required time is approximately 4 hours.
●Nearly the same discriminability as the pulsed ﬁeld gel electrophoresis（PFGE）
and Multilocus Sequence Typing（MLST）.
●This POT kit can compare the homogeny between strains by digitizing (POT type）
an analysis result.
Composition（50 rxns）

Work FlowReagentVolumeA

AptaTaq DNA Master(5×Conc.)*1500 μL×1

B

PCR supplement

500 μL×1

C

Primer mix α

250 μL×1

D

Primer mix β

250 μL×1

E

Positive control

500 μL×1

F

Loading buﬀer（6×Conc.）

500 μL×1Isolate

DNA Extraction*2

Multiplex PCR
Agarose gelelectrophoresis

comparethe homogeny*1 AptaTaq DNA Master（5×Conc.）is the product of Roche Diagnostics K.K.
*2 CicaGeneus® DNA extraction reagent is sold separately.

Kanto Chemical

Convert toPOT type.

Analyzebanding patterns

Table.1 Kind of detection ORF and Amplicon size

Reaction
mixture 1Fig.1 Example of the electrophoretic pattern
50: 50 bp DNA Ladder, P : Positive control

1: ATCCTM 27853

2: ATCCTM 35554

Reaction
mixture 2

3: PAO1

4: Clinical isolate of the IMP positive5: Clinical isolate of the VIM positive6: Clinical isolate

7,8: Clinical isolate from the outbreaks example1

9,10 : Clinical isolate from the outbreaks example2

POT No.

Amplicon size（bp）

PCR PC

506

POT 1-1

336

Genomic Islet-1

POT 1-2

281

Genomic Islet-2

POT 1-3

235

Genomic Islet-3

POT 1-4

201

Genomic Islet-4

POT 1-5

175

Genomic Islet-5

POT 2-1

151

VIM

POT 2-2

126

Propharge-1

POT 2-3103

Propharge-2

POT 2-485

Propharge-3

PCR PC506

POT 1-6324

Genomic Islet-6

POT 1-7271

Genomic Islet-7

POT 1-8

238

Genomic Islet-8

POT 1-9204

Genomic Islet-9

POT 1-10

176

Genomic Islet-10

POT 2-5150

Propharge-4

POT 2-6124

Propharge-5

POT 2-7

105

IMP

Target domain
positive control

positive control① Read banding patterns（Table.1）from an electrophoretic result（Fig.1）.
② Using a result of having this electrophoretic band or not, you input it into a calculation sheet for POT type analysis by the binary scaleand calculate a POT type of three categories.
ⅰ Whenpositive contorol is positive, the sample can confirm that it isand In the case of MDRP, the valuesof POT2 become more than 64 or odd number.
ⅱ You can suppose the homogeny between strains objectively by comparing the POT type.
ⅲ As for the strains obtained from outbreaks, all POT types（1-3 ）become same.（7&8、9&10 in Fig.1）

ResultSample number in Fig.1

Category

Product Information
Product No.

Product Name

Package sizeStored at08187-96

CicaGeneus

Pseudo POT KIT

50 rxns-20 ℃〜-25 ℃

08178-96

CicaGeneus

DNA Extraction Reagent

120 rxns

2 ℃〜8 ℃

TMTM

Multiplex PCR kit series（stored at -20 ℃ 〜 -25 ℃）
Product Name

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AptaTaq DNA Master（5×Conc.）

500 μL

1L
10 mL＊希望価格は税抜きです。

● The example data of the electrophoretic pattern is provided by Dr.Suzuki of Aichi Prefectural Institute of Public Health.
● This product is obtaining the patent licensing from Aichi Prefecture.
● The performance evaluation had Dr. Nagao of Kyoto University graduate school and Dr.Kirikae of National Center for Global Health and Medicine.

KANTO CHEMICAL CO., INC.
REAGENT DIVISION

East Muromachi Mitsui BLDG, 2-1, Nihonbashi Muromachi 2-chome,Chuo-ku, Tokyo, 103-0022, JAPAN
Telephone+81-3-6214-1092
Telefax+81-3-3241-1053
http://www.kanto.co.jp
E-mail; kanto-61@gms.kanto.co.jp
2015. August.