Cica Geneus Acineto POT KIT®

The spread of carbapenem-resistant
has become a global problem.
The POT method can not only identify the species of Carbapenem-resistant(ACB) complexes but also simultaneously determine
the international epidemic clone I, II and the genetic identities ofin several hours.
This POT method can easily and rapidly identify the international epidemic clones andthe identities of(、、、genomic species close to 13TU). It is a useful tool for infection control.
※ This test method is developed by grants from the Ministry of Health, Labour, and Welfare of Japan (grants H24-Shinko-Ippan-10)

Characteristics● This POT method developed by Dr. Suzuki of Fujita health University, it s a molecularepidemiological method which can identify and distinguish isolates with strain-level.
● This POT method can compare the homogeny between strains by digitizing(replace to POT types) an electrocataphoresis result.
● Nearly the same discriminablity as the as Multilocus Sequence typing (MLST).
● Required for the examination time is approximately 4 hours.
Composition（30 rxns）
Reagent

Work FlowVolumeIsolateA

AptaTaq DNA Master(5×Conc.)*1

240 μL×1

B

PCR supplement

240 μL×1

C

Primer mix α120 μL×1

D

Primer mix β

120 μL×1

E

Positive control

240 μL×1Analyze banding patterns

FLoading buﬀer（6×Conc.）

240 μL×1

Convert to POT type

* AptaTaq DNA Master（5×Conc.）is the product of Roche Diagnostics K.K.

DNA Extraction*2Multiplex PCR
Agarose gel electrophoresis

1*2 CicaGeneus® DNA extraction reagent is sold separately.

Kanto Chemical

Compare the homogeny

Table.1 Kinds of detection ORF and Amplicon size
POT No.
PCR PCAmplicon size (bp) POT modulus Target domain
atpA
553
A. baumannii
465
OXA-51401
1000 Speciﬁc gene
A. pittii
A. nosocomialis
362
2000 Speciﬁc geneA. sp. close to 13TU
321
3000 Speciﬁc gene
Reaction
POT 1-1
271
64
Genomic Islet-1
mixture 1
POT 1-2
231
32
Genomic Islet-2
POT 1-3
189
16
Genomic Islet-3
POT 1-4
151
8
Genomic Islet-4
POT 1-5
122
4
Genomic Islet-5
POT 1-6
102
2
Genomic Islet-6
POT 1-7
81
1
Genomic Islet-7
PCR PC565
OXA-51
POT 2-1
457
32
Genomic Island-1
POT 2-238816
Genomic Island-2
POT 2-3329
8
Genomic Island-3
POT 2-4280
4Genomic Island-4
POT 2-5
237
2
Genomic Island-5
Reaction
POT 2-6
209
1
Genomic Island-6
mixture 2
POT 3-1
185
32
Genomic Island-7
POT 3-2
160
16
Genomic Island-8
POT 3-3
139
8
Genomic Island-9
POT 3-4120
4Genomic Island-10
POT 3-5
101
2
Genomic Island-11
POT 3-6
81
1
Genomic Island-12

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

1,2：
3：
4：
5：
6：
7：
8：
9：
10：
11：

（ST2）(Clinical isolate from the outbreaks example)（ST2）（ST2）Multiple-Drug-Resistant（ST1）ATCC™ BAA1605（ST1）（ST152）（ST34）

sp. close to① Distribution patterns of ORFs are identiﬁed by reading electrophoresis banding patterns（Fig.1）.
② POT types of three categories are obtained by input electrophoresis banding patterns into the Excel calculation sheet
which can be downloaded from Kanto Chemical Co. Inc web site.
ⅰ When PCR PC is positive, the sample can confirm that it isⅱ The value of POT 1 ofbecome less than 1000,
become 1000-1999,
sp. close to 13TU become 3000-3999, Otherspp. bocome more than 4000.
ⅲ If the value of POT 1 become 69, the strain is supposed to international clone Ⅰ,
If it become 122, the strain is supposed to international clone Ⅱ,
ⅳ You can suppose the homogeny between strains objectively by comparing the POT type.

become 2000-2999,

ⅴ As for the strains obtained from outbreaks, all POT types（1-3 ）become same.（1&2 in Fig.1）

ResultCategory
POT1
POT2
POT3

1
122
59
54

212259
54

3122
2654

Sample number in Fig.1

569
4659

412253
4166942277720891105
00810480113105
0010210500■ Product Information
Product No.

Product Name

Package size

Stored at08０６２-96

CicaGeneus® Acineto POT KIT

30 rxns-20 ℃〜-25 ℃

08178-96

CicaGeneus® DNA Extraction Reagent

120 rxns

2 ℃〜8 ℃

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

Product Name

Related reagents

Package size

Product No.

Product Name

Package size
1L08180-96

Cica Geneus® Staph POT KIT

120 rxns

46510-79

10×TBE buﬀer

08187-96

Cica Geneus® Pseudo POT KIT

50 rxns

46509-79

10×TAE buﬀer

08362-97

Cica Geneus® E.coli POT KIT

30 rxns

14575-43

Ethidium bromide solution

08106-97

Cica Geneus® C. diﬀ POT KIT

30 rxns

01089-23

Agarose KANTO HC

100 g

08143-96

Cica Geneus® AmpC Genotype Detection KIT

30 rxns

01016-96

AptaTaq DNA Master（5×Conc.）

500 μL08112-96

Cica Geneus® ESBL Genotype Detection KIT

30 rxns

08158-96

Cica Geneus® Carbapenemase Genotype Detection KIT2

30 rxns

1L
10 mL● The example data of the electrophoretic pattern is provided by Dr. Arakawa of Nagoya University and Dr.Suzuki of Fujita Health University.
● This product is obtaining the patent licensing from Aichi Prefecture and Nagoya University.

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
2018 OCT.