Cica Geneus Carbapenemase
Genotype Detection KIT2®

Carbapenemase Genotype Detection
IMP-1, VIM, KPC, NDM, OXA-48 group+ GES group (Carbapenemase type)+ IMP-6 gene

Carbapenem exhibits antibacterial activities and a very wide antibacterial spectrum.
Recently, increasing of Carbapenem Resistant Enterobacteriaceae (CRE) becomes
global problem. And stealth-type CRE, which is diﬃcult to detect by conventional
drug sensitivity test, exists in Carbapenemase Producing Enterobacteriaceae (CPE).
This kit can detect 6 representative Carbapenemase genotypes anddistinguish IMP-6 gene from IMP-1 group by two kinds of multiplex PCR.
This kit was collaborativery deveroped by Prof. Dr. Sugai and Assistant Prof. Dr.
Kayama at Hiroshima University.

Characteristics● This kit can detect 6 carbapenemase genotype(IMP-1 group, VIM group, KPC group, NDM group, OXA-48 group,GES group(carbapenemase type), and distinguish IMP-6 gene from IMP-1.)● Required time is approximately 3 hours and suitable for use in common thermal cycler.

Work Flow

Composition（30 rxns）
Reagent

VolumeIsolate carbapenemase producing bacteria*2A

AptaTaq DNA Master(5×Conc.)

240 μL×1

B

PCR supplement

420 μL×1

C

Primer mix α

120 μL×1

D

Primer mix β

120 μL×1

Agarose gel electrophoresis

E

Positive control

100 μL×1Detection

FLoading buﬀer（6×Conc., Orange-G）

240 μL×1*1*1 AptaTaq DNA Master（5×Conc.）is the product of Roche Diagnostics K.K.

DNA Extraction*3
Multiplex PCR*2 CHROMagar™ mSuperCARBA (72141) is recommended.
*3 Cica Geneus® DNA extraction reagent is sold separately.

操作手順

Table1 Target genes and its amplicon size
Target gene
Reaction mixture.1

Reaction mixture.2

Amplicon size（bp）
587
VIM group
401
GES(Carbapenemase) group
228
KPC group798
NDM group616
OXA-48 group
442
IMP-6
282
IMP-1 group① Please refer to an attached document.
② From an electrophoretic pattern of two
multiplex PCR (Figure. 1), you can estimate
carbapenemase genotype (Table.1)
in comparison with the band of the positive control.

Fig.1 Example of the electrophoretic pattern
100：100 bp DNA Ladder, P：Positive control（Reagent E）,
1：IMP-1 positive bacteria, 2：VIM positive bacteria,
3：GES(Carbapenemase type) positive bacteria,
4：GES(ESBL type) positive bacteria
5：KPC positive bacteria, 6：NDM positive bacteria,
7：OXA-48 positive bacteria, 8：IMP-6 positive bacteria,
N：Negative control（ TE Buﬀer）● This kit can not identify bacterial species.
● Even if the carbapenemase genes are detected by this kit,
the strain may not be resistant to antibiotics because of
the gene mutations or expression eﬀects.
● Some carbapenemase genes can not be detectedby this kit. Additionally, even if the strain has certain
target genotypes, it may be undetectabledepending on subtypes.
● In this kit, GES genes with G170N or G170S mutation areexpressed as carbapenemase type GES genes,and otherwise are expressed as ESBL type GES genes.
● Use TAE buﬀer for electrophoresis.
● This kit is for research and experimental use.

● Product Information
Product No.

Product Name

Package size

Stored at

08158-96

Cica Geneus Carbapenemase Genotype Detection KIT2

30 rxns-20 ℃〜-25 ℃

08178-96

Cica Geneus DNA Extraction Reagent

120 rxns

2 ℃〜8 ℃

Package size

Stored at

120 rxns-20 ℃〜-25 ℃

30 rxns-20 ℃〜-25 ℃®

®

● Multiprex PCR kit series
Product No.
08180-96
08180-97

Product Name
Cica Geneus® Staph POT KIT

08187-96

Cica Geneus Pseudo POT KIT

50 rxns-20 ℃〜-25 ℃

08062-96

Cica Geneus® Acineto POT KIT

30 rxns-20 ℃〜-25 ℃

08362-97

Cica Geneus E.coli POT KIT

30 rxns-20 ℃〜-25 ℃

08112-96

Cica Geneus® ESBL Genotype Detection KIT

30 rxns-20 ℃〜-25 ℃

08143-96

Cica Geneus AmpC Genotype Detection KIT

30 rxns-20 ℃〜-25 ℃

®

®

®

● Related reagent
Product Name

Product No.
72141

CHROMagar™ mSuperCARBA (prepared)

Package size
10 sheets
1L46509-79

10×TAE buﬀer

14575-43

Ethidium bromide solution

10 mL

01098-23

Agarose KANTO LE

100 g

01016-96

AptaTaq DNA Master（5×Conc.）

500 μL←Please acsess Cica-web
to catch the detail ofour reagent information !

● This product is developed with Hiroshima 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
2018/05