Cica Geneus AmpC Genotype Detection KIT®

AmpC producing bacteria, which susceptibility is similar to ESBL-producing bacteria, areresistant bacteria inhibited by boronic acid, cloxacillin, etc. They are concerned aboutbecoming ineﬀective to antibacterial drugs of cephalosporin and cephamycin type.
This kit can detect 6 representative plasmid-mediated AmpC gene family by two
kinds of multiplex PCR.
This kit was collaborativery deveroped by Prof. Dr. Sugai, Assistant Prof. Dr. Kayamaand MMSc. Hara at Hiroshima University.

Characteristics● This kit can detect 6 AmpC gene family(CIT family, FOX family, ACT family, DHA family, ACC family, and MOX family).

● Required time is approximately 3 hours.
● Suitable for use in common thermal cycler.

Composition（30 rxns）
Reagent

Work Flow
VolumeA

AptaTaq DNA Master(5×Conc.)*1

240 μL×1

B

PCR supplement

420 μL×1

C

Primer mix α

120 μL×1

D

Primer mix β

120 μL×1

E

Positive control

100 μL×1

Detection

FLoading buﬀer（6×Conc., Orange-G）

240 μL×1*2 CHROMagar™ AmpC/ESBL diﬀerential discs (74054) is recommended.

* AptaTaq DNA Master（5×Conc.）is the product of Roche Diagnostics K.K.
1Isolate AmpC producing bacteria*2
DNA Extraction*3
Multiplex PCR
Agarose gel electrophoresis*3 Cica Geneus® DNA extraction reagent is sold separately.

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

CIT family
FOX family
ACT family

Reaction mixture.2

DHA family
ACC family
MOX family

Amplicon size（bp）
576
364
213
619
403
266

① Please refer to an attached document.
② From an electrophoretic pattern of twomultiplex PCR (Figure. 1), you can estimate AmpC
genotype (Table.1) in comparison with the band of
the positive control.
● This kit can not identify bacterial species.
● Even if the AmpC genes are detected by this kit,
it may not be resistant to antibiotics because of
the gene mutation or expression eﬀects.
● Some AmpC genes can not be detected by this kit.
Also, even if the strain has certain target genotypes,
it may not be detectable depending on subtypes.
● Chromosomal AmpC genes are also detected in bacterial
species that carry the certain target family ofAmpC genes on their chromosomes.
● Use TAE buﬀer for electrophoresis.
● This kit is for research and experimental use.

Fig.1 Example of the electrophoretic pattern
100：100 bp DNA Ladder, P：Positive control（Reagent E）,
1：CIT positive bacteria, 2：FOX positive bacteria,
3：ACT positive bacteria, 4：DHA positive bacteria
5：DHA positive bacteria, 6：MOX artiﬁcial gene,
N：Negative control（ TE Buﬀer）● Product Information
Product No.

Product Name

Package size

Stored at08143-96

Cica Geneus AmpC Genotype Detection KIT

30 rxns-20 ℃〜-25 ℃

08178-96

Cica Geneus DNA Extraction Reagent

120 rxns

2 ℃〜8 ℃®

®

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

Product Name
Cica Geneus® Staph POT KIT

Package size

Stored at

120 rxns-20 ℃〜-25 ℃

30 rxns-20 ℃〜-25 ℃

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 ℃

08158-96

Cica Geneus® Carbapenemase Genotype Detection KIT2

30 rxns-20 ℃〜-25 ℃● Related reagent
Product Name

Product No.
72141

CHROMagar™ mSuperCARBA (prepared)

Package size
10 sheets
1L

46509-79

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.