Cica Geneus Staph POT KIT
TM

Methicillin-resistant（MRSA）is pathogens of the hospital-acquired infections, and it is necessary to take appropriate diﬀusion precaution.
The PCR-based ORF Typing（POT）*1 developed by Dr.Suzuki of Aichi Prefectural Institute of Public Health is a method to amplify plural speciﬁc genes by Multiplex PCR at
the same time, and then to analyze agarose gel electrophoresis banding patterns.
References
1. Suzuki.M et.al.
2. Suzuki.M et.al.

101(4):938-947 (2006)
107(4):1367-1374 (2009)

Characteristics● Required time is approximately 4 hours.
● Nearly the same discriminability as the pulsed ﬁeld gel electrophoresis（PFGE）.
● This POT kit can compare the homogeny between strains by digitizing (POT type）
an analysis result.
■ Work Flow

■ Composition（120 rxns）
Reagent

VolumeA

AptaTaq DNA Master(5×Conc.)

B

PCR supplement

1.0 mL×1

C

Primer mix α

0.5 mL×1

D

Primer mix β

0.5 mL×1

E

Positive control

1.0 mL×1

F

Loading buﬀer（6×Conc.）

1.0 mL×1*1Isolate

1.0 mL×1

DNA Extraction*2

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

Kanto ChemicalConvert to
POT typeAnalyze
banding patterns

Table.1 Kind of detection ORF and Amplicon size
POT No.

Amplicon size（bp）

Target domain
positive control

601

Reaction
mixture 1

POT 1-1

530

POT 1-2

449

POT 1-3

355

SCC

POT 2-1

304

Tn

POT 2-2271

Propharge

POT 2-3

228

Propharge

POT 2-4

197

Propharge

POT 2-5161

Propharge

POT 2-6131

Propharge

POT 2-7106Propharge

POT 2-881

Genomic Islandgene complex class B
type Ⅱa speciﬁc

positive control

601

Fig.1 Example of the electrophoretic patternｓ
50 : 50 bp DNA Ladder, P : Positive control
1: ATCCTM 700699（MRSA Mu50）

2: ATCCTM BAA-1556（MRSA USA300）

3: ATCCTM 43300（MRSA）

4: ATCC

5: ATCCTM 29213（MSSA）

TM

Reaction
mixture 225923（MSSA）

6,7 : Clinical isolate from the outbreaks example1
8,9 : Clinical isolate from the outbreaks example2

POT 1-4477

cassette chromosome recombinase A2

POT 1-5

388

Genetic back groundPOT 1-6

320

Genetic back groundPOT 1-7

273

POT 3-1

243

Propharge

POT 3-2

197

Propharge

POT 3-3

171

Propharge

POT 3-4140

Propharge

POT 3-5115

Propharge

POT 3-695

Propharge

POT 3-778

Propharge

gene complex class A① 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 Whenis positive, the sample can confirm that it isand In the case of MRSA, the values of POT1

become more than 64.
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.（６&７、８&９ in Fig.1）

ResultSample number in Fig.1

Category

Product Information
Product No.

Product Name

Package size

Sｔｏｒｅd at

08180-96

CicaGeneus

Staph POT KIT

120 rxns-20 ℃〜-25 ℃

08178-96

CicaGeneus

DNA Extraction Reagent

120 rxns

2 ℃〜8 ℃

TMTM

Multiplex PCR kit series（stored at -20 ℃ 〜 -25 ℃）
Product No.

Product Name

Package size

Related reagents
Product Name

Product No.

Package size

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50 rxns

46510-79 10×TBE buﬀer

08062-96 CicaGeneusTM Acineto POT KIT

30 rxns

46509-79 10×TAE buﬀer

08087-96 CicaGeneusTM Pathogenesis Gene Detection PCR Kit（for DEC）

100 rxns

14575-43 Ethidium bromide solution

08179-96 CicaGeneusTM Staph coaglase detection set

50 rxns

01089-23 Agarose KANTO HC

100 g

08081-96 CicaGeneusTM Bovine Mycoplasma HI screening kit

100 rxns

01016-96 AptaTaq DNA Master（5×Conc.）

500 μL

1L1L
10 mL●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.

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.