施設数 割合(%) 施設数 割合(%) 施設数 割合(%)

t r

1 8MERS, SFTS 8

e

B B8 e S 8

d 8 28 e d

8 B

2 AMR CRE

b 8 8

d 8CRE E

b

1 8MERS, SFTS

8

e

B B8

e B D8

b8 b

2 AMR CRE

b 8

1.

e 8

8

2. E CRE

E CRE 8 B

8 B

1.

8 u A H1N1

pdm098 uH7N98MERS,

SFTS 8

8 b8

e B

8 e e 8 B

B B

28 4

- 11 -

B 8 D

B 8

B be

d S 28 e8

d 8

8 B

2. E CRE

CRE 8 E

B

D8 IMP PCR

B e

D i e 8

B 8

8 8

BS D b8

b c B

8CRE

b 8 8 d 8

E CRE b

1

○

○ (MEPM) 8

(IPM) (CMZ)

2

○

IMP 8NDM 8KPC 8OXA-48 4

○ β- r (ESBL)

TEM 8SHV 8CTX-M-1 group8 CTX-M-2 group8CTX-M-9 group 5

○AmpC β- r

MOX 8CIT 8DHA 8ACC 8EBC 8FOX 6

↓

c 8

SMB 8KHM 8VIM 8GES 8IMI

3 β- r

○r ESBL

○ KPC

○ r tv

AmpC

○ (SMA) β-

r (MBL)

4 MBL

○Carba NP

○Carbapenemase Inactivation Method (CIM )

*2 3,4

B

5 l s (PFGE)

d B i

6 NGSB

- 12 -

S1-PFGE NGS

1 2 CRE

CRE

AMR

1．Doan YH, Haga K, Fujimoto A, Fujii Y, Takai-Todaka R, Oka T, Kimura H, Yoshizumi S, Shigemoto N, Okamoto -Nakagawa R, Shirabe K, Shinomiya H, Sakon N, Katayama K. Genetic analysis of human rotavirus C: The appearance of Indian-Bangladeshi strain in Far East

Asian countries. Infect Genet Evol.

41:160-73. 2016

2. Suzuki Y, Doan YH, Kimura H, Shinomiya H, Shirabe K, Katayama K.

Predicting genotype compositions in norovirus seasons in Japan. Microbiol Immunol. ;60(6):418-26. 2016

3. Okamoto K, Mori Y, Komagome R, Nagano H, Miyoshi M, Okano M, Aoki Y, Ogura A, Hotta C, Ogawa T, Saikusa M, Kodama H, Yasui Y, Minagawa H,

Kurata T, Kanbayashi D, Kase T, Murata S, Shirabe K, Hamasaki M, Kato T, Otsuki N, Sakata M, Komase K, Takeda M. Evaluation of sensitivity of TaqMan RT-PCR for rubella virus detection in clinical specimens. J Clin Virol. ;80:98-101. 2016,

4. Kobayashi M, Matsushima Y, Motoya T, Sakon N, Shigemoto N, Okamoto- Nakagawa R, Nishimura K, Yamashita Y, Kuroda M, Saruki N, Ryo A, Saraya T, Morita Y, Shirabe K, Ishikawa M, Takahashi T, Shinomiya H, Okabe N, Nagasawa K, Suzuki Y, Katayama K, Kimura H. Molecular evolution of the capsid gene in human norovirus genogroup II. Sci Rep. Jul 7;6:29400.

2016

5

D68

. 44(4) 156-159 2016 6

- 13 -

8 846 12 810−1182017,

1. 8 d

n 8

n 8

l v

t 7 25 6 7 011 2-

8 8

a

- 14 -

- 15 -

- 16 -

- 17 -

28

DNA

enterohemorrhagic E. coli: EHEC 2016 3,640

1,250

[hemolytic uremic syndrome: HUS]

NESID

O157 O26, O111, O103, O145, O121, O165

EHEC

EHEC EHEC

[enteropathogenic E. coli: EPEC]

[enteroaggregative E. coli: EAggEC]

Escherichia

albertii PCR

15 1

8 2

19 8

Legionella pneumophila

- 18 -

A

A Group A

Streptococcus Streptococcus pyogenes A

A

A

A A

A

A

EHEC

A A

1.

Staten Serum Institut: SSI

PCR

2. SBT

L. pneumophila EWGLI

(European Working Group of Legionella Infections) SBT (sequence-based

typing) flaA pilE asd mip

mompS proA neuA

(http://www.hpa-bioinformatics.org.uk/legi onella/legionella_sbt/php/sbt_homepage.php)

3. T M

1.1 EHEC

- 19 -

2016

EHEC 2,676

O157 57.8% O26 24.4%

O103 4.5% O111 2.7% O121 1.9%

O145 1.5% O91 1.2% O165 0.3%

5.7%

1.2

EHEC, EPEC, EAggEC, ETEC [enterotoxigenic

E. coli: ], EIEC

[enteoinvasive E. coli:

] EHEC

DNA

1.3 Escherichia albertii

E. albertii PCR

1.4 EQA External

Quality Assurance

Statens

Serum Institut: SSI

EQA

2016 10

EHEC

EQA

, β ,

O:H

SSI 1.5 EHEC

2.

53

Legionella pneumophila SG

1 48 SG2 9 13 1

Legionella longbeachae SG2 Legionella bozemanae SG2 1

2015

2 1 SG1 SG13

1 2

PFGE

PFGE 1

DNA

- 20 -

3 1

3

1 3 1

2016 3 436

452 L. pneumophila 426 97.7%

L. pneumophila 1 85%

L. pneumophila

426

ST1 ST2134 187

STSS 110 emm1

41 (37.3%) emm89

27 (24.5%) emm12 9 (8.2%)

2014 emm1

emm89 16.0% (12/75) 24.5% (27/110)

- 21 -

EQA

1) Ikebe T, Matsumura T, Nihonmatsu H, Ohya H, Okuno R, Mitsui C, Kawahara R, Kameyama M, Sasaki M, Shimada N, Ato M, Ohnishi M. Spontaneous mutations in Streptococcus pyogenes isolates from streptococcal toxic shock syndrome patients play roles in virulence. Sci Rep 6:28761, 2016.

2) Kuroki T, Amemura-Maekawa J, Ohya H, Furukawa I, Suzuki M, Masaoka T, Aikawa K, Hibi K, Morita M, Lee K, Ohnishi M,

- 22 -

Kura F. Outbreak of Legionnaire’s disease caused by Legionella pneumophila serogroups 1 and 13. Emerg Infect Dis. 23 349-351, 2017.

3)

1 Amemura-Maekawa J, Chida K, Ohya H,

Isobe J, Kanatani J, Tanaka S, Nakajima H, Yoshino S, Ohnishi M and Kura F:

Characterization for clinical Legionella species by Legionella Reference Center in Japan. ESGLI 2016. Amsterdam, September 2016.

- 23 -

- 24 -

- 25 -

- 26 -

- 27 -

- 28 -

- 29 -

- 30 -

- 31 -

- 32 -

- 33 -

- 34 -

2016

PCR

( )

QOL

BSL3

1.

PCR

- 35 -

( )

2. Probe

PCR

3.

1. PCR

6

PCR PCR

2. Probe

3.

PCR

R. japonica

R.

heilongjiangensis R.

tamurae R. helvetica

Karp Kato Gilliam Kawasaki Kuroki

Shimokoshi

- 36 -

PCR

PCR

1. Satoh M, Akashi S, Ogawa M, Wakeyama T, Ogawa H, Fukuma A, Taniguchi S, Tani H, Kurosu T, Fukushi S, Shimojima M, Ando S, Saijo M: Retrospective survey of severe fever with thrombocytopenia syndrome in patients with suspected rickettsiosis in Japan. J. Infect.

Chemother., 23: 34-50, 2017 2.

2011 2014

, 67(3): 167-175, 2016

1.

28

2017 2 21 22 2.

update

23 2016 12 3 4

- 37 -

- 38 -

- 39 -

- 40 -

- 41 -

- 42 -

, ,

, , , ,

, , ,

27

- 43 -

- 44 -

- 45 -

- 46 -

- 47 -

- 48 -

- 49 -

Loopamp D

Loopamp

D LAMP

ptxP1

ptxP3 SNP; G>A at −65 position ptxP8 C>T at −60 position

ptxP1 ptxP3 99

2016 11 Loopamp

D LAMP

LAMP

ptxP1 ptxP SNP

ptxP1 ptxP3

ptxP1 ptxP3 ptxP8

ptxP

SNP LAMP

LAMP ptxP3

ptxP8

1. ptxP DNA

ptxP1 ptxP3

ptxP8 PCR 3

ptxP ptxP1, ptxP3, ptxP8

ptxP 341-bp QIAEX II Gel Extraction Kit

ptxP1 n=20 ptxP3 n=20 ptxP8

n=1 1 Wizard

Genomic DNA Purification Kit DNA

2. Loopamp D

RNA (0.1 g/mL) TE ptxP

10⁵ 10² DNA

DNA 100 pg 1 pg DNA

DNA Loopamp

- 50 -

D LMP542 LA-320C ptxP

66 40 Tt OD650

0.1 min

1. ptxP

PCR 3 ptxP

ptxP1, ptxP3, ptxP8 10² 10⁵

ptxP1 ptxP8 Tt

ptxP3 ptxP1 ptxP8

Tt p < 0.05 10³~10⁵

ptxP3 ptxP1 ptxP8

100

2. DNA

ptxP

ptxP1 ptxP8 Tt

ptxP3 ptxP1 Tt p < 0.01 DNA

ptxP3

LAMP

ptxP1 ptxP3 ptxP8

ptxP3 LAMP SNP

ptxP1 ptxP8

LAMP 2~3

SNP

ptxP3 SNP DNA LAMP

ptxP ptxP1 ptxP3

99 LAMP

ptxP1 ptxP3

ptxP

ptxP

Loopamp D

ptxP1 ptxP3

1. Hiramatsu Y, Miyaji Y, Otsuka N, Arakawa Y, Shibayama K, Kamachi K. Significant decrease in pertactin-deficient Bordetella pertussis isolates, Japan. Emrg Infect Dis. in press.

2. Kamachi K, Moriuchi T, Hiramatsu Y, Otsuka N, Shibayama K. Evaluation of a commercial loop-mediated isothermal amplification assay for

- 51 -

diagnosis of Bordetella pertussis infection. J Microbiol Methods.

133:20-22, 2017.

1.

48

11 19-20

2016

Fig. 1. Schematic representation of the LAMP primers for Bordetella pertussis detection.

The LAMP primers are designed to target the B. pertussis ptxP1 allele. The ptxP3 and ptxP8 alleles contain a SNP at positions -65 (G>A) and -60 (C>T), respectively. The transcription start site is shown at position +1. ptxA, pertussis toxin S1 subunit gene.

Fig. 2. Standard curves for ptxP1, ptxP3, and ptxP8 alleles generated with PCR fragments.

The LAMP assay was performed with 10-fold serial dilutions of PCR fragments (10⁵ to 10² copies per reaction). Data represent the mean and standard deviations from three separate measurements.

- 52 -

Table 1. Detection of Bordetella pertussis ptxP1, ptxP3, and ptxP8 strains by the commercial LAMP assay

Strain No. of clinical strains tested^a

Tt value (min)^b

100 pg DNA^c 1 pg DNA

ptxP1 20 19.3 ± 0.5 21.8 ± 0.5

ptxP3 20 16.9 ± 0.3 19.0 ± 0.4

ptxP8 1 18.5^d 21.5^d

a Genomic DNA sample prepared from B. pertussis clinical isolate.

b Threshold time when the turbidity value reached OD650 of 0.1.

c Genomic DNA/reaction tube.

d Average Tt values from two separate measurements.

- 53 -

VNTR

VNTR

2014 VNTR External

Quality Assessment: EQA

2016

2014 2015

3

2016 56 55

3 JATA 12

12 48 87%, 48/55

2016 2015 93%, 49/53

2014 67%, 36/54 P=0.01

VNTR Variable Number of Tandem Repeat

VNTR

2014 VNTR 3 JATA 12 -VNTR

66.7%

36/54

2016

2014

2015 3

Quality Assurance: QA

Internal Quality Control: IQC External Quality Assessment: EQA

Training: TA

- 54 -

VNTR

DNA

DNA 3 3

DNA 2

DNA DNA

DNA

VNTR VNTR

PCR

: JATA 12 JATA 15 Supply 15

HV Hypervariable Regions/ : 3232, 3820, 4120

JATA 12

VNTR

PCR VNTR

1.

79

2016

11 2016 12 56

2017 3

1 1 55

55

2. VNTR

VNTR JATA 12

JATA 15 HV Supply[15]

JATA

12 JATA

15 JATA[12] 3 HV

3 Supply 6

2016

JATA 15 HV Supply 41 33 19

2014 2015

1

3. JATA 12

3

JATA 12 12

48 87%,

- 55 -

48/55 1 5 9.1%,

5/55 2 2 3.6%,

2/55 1 2016

2014

67% vs. 87%, p=0. 01 2015

93% vs.87%, p=0.24

2016 2015

1

1.9% vs. 9.1%

4. PCR PCR

QIAxcel QIAGEN

2 2016 2015

66%, 36/55

10 18%, 10/55 5

9%, 5/55 QIAxcel 2 4%, 2/55 Agilent 2100 Bioanalyzer 1 2%, 1/55

5.

PCR

JATA 12 JATA 15 HV Supply 3

1

2016 2014

2015

JATA 12

2016 2015

99.7% [2015] vs. 99.8% [2016]

10

100% [2015] vs. 98.9% [2016]

5

100% [2015] vs. 97.2% [2016]

JATA 15 Supply

99.6%-100%

HV

97.2%

[2015] 97.8% [2016]

100% [2015] vs 98.8% [2016]

1

Agilent 2100 Bioanalyzer QIAxcel

6.

JATA 12 JATA 15

3 2014 5 1955,

3336, 4052, 4156, 2163a 77–96% 2015

99–100%

2016 98-100%

: 5

- 56 -

[2015] vs. 9 [2016]

2016

2014

2015 3

PCR

2016 2014

2015 2

36

10

5 QIAxcel 2 Agilent 2100 Bioanalyzer 1

3 JATA 12

2016 2014

67% vs. 87%, p=0. 01 2015 93% vs.87%,

p=0.24 2016

2015 1

1 vs. 5 1

4

data not shown

VNTR DNA

7 5

VNTR

data not shown

2016 2014

2014 5

2015 2016

2

2015 VNTR DNA 2016

VNTR DNA

VNTR

JATA 15 HV Supply 6

41 33 21 3

2 JATA 12

JATA 12

- 57 -

2014 2015 2016 VNTR

VNTR

VNTR

2016 55 VNTR

3 JATA

12 12

46 87%, 48/55 2016

2015 93%, 49/53

2014 67%, 36/54 VNTR

1.

.

2014-2015 . 92

. 3 23-24 , 2017 , .

- 58 -

2014 施設数 （54施設中, %）

2015 施設数 （53施設中, %）

2016 施設数 （55施設中, %）

全ローサイ完全一致 36 　67％（36/54） 49 　93％（49/53） 48 　87％（48/55）

１ローカス違い 7 　13％（7/54） 1 　 1.9％（1/53） 5 　9.1％（5/55）

２カ所以上違い 11　 20％（11/54） 3 　 5.7％（3/53） 2 　3.6％（2/55）

1. 3 JATA(12)

施設数 割合(%) 施設数 割合(%) 施設数 割合(%)

アガロースゲル 37 69 34 64 36 66

自動シーケンサー 7 13 10 19 10 18

マルチナ 4 7.4 4 7.5 5 9.1

QIAxcel 4 7.4 3 5.7 2 3.6

コスモアイ 2 3.7 2 3.8 1 1.8

Agilent 2100 Bioanalyzer

1 1.8

2014

2. PCR

- 59 -

n ^正答率(%) n ^正答率(%) n ^正答率(%) n ^正答率(%) アガロースゲル 37 98.5 22 94.4 15 94.8 5 96.7

自動シーケンサー 7 97.6 7 92.1 7 92.1 7 95.2

マルチナ 4 96.5 2 83.3

QIAxcel 4 86.1 4 80.6 4 75 1 94.4

コスモアイ 2 98.6 2 83.3 1 100 1 100

アガロースゲル 34 99.7 22 100 16 97.2 5 100

自動シーケンサー 10 100 9 100 10 100 9 100

マルチナ 4 100 2 100 2 100 1 100

QIAxcel 3 99.1 2 94.4 2 66.7

コスモアイ 2 100 1 100 1 100

アガロースゲル 36 99.8 27 99.6 20 97.8 8 100

自動シーケンサー 10 98.9 9 100 9 98.8 9 100

マルチナ 5 97.8 3 100 2 100 1 100

QIAxcel 2 97.2 1 88.9 1 66.7

コスモアイ 1 100 1 100 1 100 1 100

Agilent 2100 Bioanalyzer 1 100 2014

2015

JATA(12) JATA(15) HV Supply

2016

3.

n:

(%): 1 (%)

- 60 -

28

conventional PCR EQA 24

3 SOP

1,000 23

10pg/µl

7

7 16

PCR

SFTS

ELISA

EQA 7

14 SFTS

7

EQA

1,400

2008 7 4

16srRNA

fopA PCR

- 61 -

7

EQA 17

24 EQA

1.

Yama

3 EQA

SOP

3 No.1 2 3

1 33 1 3 1 15

7 1

96

SOP 3 3

2. conventional PCR 16srRNA fopA

LVS

100pg/µl 3 (

100pg/µl)

SOP EQA

3 No.1 2 3 Francisella novicida U112 Francisella philomiragia 029 Wolbachia sp.

SOP

10 PCR

3

PCR 16srRNA

fopA

SOP

1.

OD560

11 0.94 1.3

24 96

16

320 640

40 80 10

1 No.1

10 1 10 12 20

10 40 1

No.2 80 18 160

6 No.3 20 13

40 11 2

SOP

- 62 -

2. conventional PCR

24 16srRNA

PCR 1pg-1fg/µl

10^-2 10^-5 fopA

10pg-10fg/µl 10^-1 10^-4

3 16srRNA

fopA PCR

No.1 +/+ No.2 +/- 24

No.3 23

-/- 1 -/+ 1

No.3 -/+

Fast PCR

SOP 2 PCR

6

Takara ExTaq HS

ExTaq 11 10

PCR 4

SOP

PCR

EQA SOP

No.1 10 40

40 1 96

EQA

80

No.2 4

No.3

PCR 24

1,000

10pg/µl 600 copies/µl

EQA

Francisella

Wolbachia No.3

fopA PCR

PCR

Fast PCR

No.3

16srRNA

fopA PCR +/+

F. tularensis F. novicida +/- F. tularensis F. novicida

- 63 -

Francisella -/- Francisella -/+

EQA PCR

SOP PCR

Francisella

EQA

24

SOP

1.

1.

- 64 -

1

10 1

- 65 -

2 3

No.1 No.2 3 1

- 66 -

3 PCR 16srRNA fopA

PCR 1,000

- 67 -

HIV

HIV

HIV

HIV

NESID

2006

1500 HIV

3 AIDS

HIV

HIV

HIV

AIDS 1000 HIV

500 HIV

HIV

2

1

HIV HIV

HIV

- 68 -

2. HIV

IASR

HIV

HIV

2

HIV HIV

3

2014 10

AIDS

HIV

HIV

1 Nii-Trebi N, Ishikawa K, Matsuoka S, Takeda S, Bonney E.Y, Ofori S.B., Yoshimura K., Ampofo W.K. and Matano T. Analysis of HLA genotypes in HIV-1-infected Ghanaians. ASLM 2016 Conference. December 3-6, 2016. Cape Town, South Africa.

2 Matsuoka S. HI-1 epidemiology in Ghana.

7^th KOREA-JAPAN joint symposium on HIV/AIDS. January 14 2017. Soul, South Korea.

3

- 69 -

D B 52

54 2015 2016

79

2015 EKC

54 2

54

1. EKC 2013 2016

EKC

2017 3

EKC

803

2.

24 2012 2

5 3 EKC

3.

19a 64

- 70 -

1. EKC

D 58 B 3 18 E

4 11 13

C

2 5

D EKC

54 24

37 56 8 19 53

1

2.

2017 3

EKC 79

3.

NESID

48 49 55 57

64 19a

(2017 2 9 )

EKC adenovirus

2013 2014 2015 2016

Ade 54 12 10 91 80

Ade 3 18 48 41 37

Ade 37 10 56 34 10

Ade 4 35 23 19 9

Ade 56 19 29 14 11

Ade 8 17 13 8 2

Ade 19 1 4 4 21

Ade 53 3 4 8 6

Other

adeno 23 21 25 37

54

2015

1

2015 2016 2

54 EKC

EKC D 54

6 54 37 56 8

19 53

B 3 E 4 EKC

EKC

54

EKC

19 EKC

19a

EKC 19a

64

- 71 -

NESID 64 64 (19a)

EKC

(53 )

1. Yoshitomi H, Sera N, Gonzalez G, Hanaoka N, Fujimoto T. First isolation of a new type of human virus (genotype 79), species Human mastadenovirus B (B2) from sewage water in Japan. J Med Virol. 2016 Dec 12. doi: 10.1002/jmv.24749.

2. Hai le T, Thach HN, Tuan TA, Nam DH, Dien TM, Sato Y, Kumasaka T, Suzuki T, Hanaoka N, Fujimoto T, Katano H, Hasegawa H, Kawachi S, Nakajima N. Adenovirus Type 7 Pneumonia in Children Who Died from Measles-Associated Pneumonia, Hanoi, Vietnam, 2014. Emerg Infect Dis.;22(4):687-90, 2016.

3.

-2014 - 90(4) 507 511 . 2016 4.

27(1) 69 73 2016

5. 460 11,

2016.

6. 461

11,2016

7. .

462 11, 2016

8. .

87(6) 748 750 2016

1 Hanaoka N. Adenovirus Infection as STI; adenoviral urethritis. 19th IUSTI Asia-Pacific Conference. Dec-2, 2016, Okayama city

2.

(2014 ) 53 7 2 , 2016 , 3.

.

17 7 2

2016 4.

37

7 22 2016

5.

53 54 56

2000 2015

90 4

15 2016

6.

- 72 -

90 4

16 2016

7. Aksara Thongprachum, Tsuguto Fujimoto, Sayaka Takanashi, Shoko Okitsu, Satoshi Hayakawa, Hiroshi Ushijima. A variety of virus commonly causing diarrhea detected in untreated sewage. The 64^th annual meeting of the Japanese Society of Virology. October 25^th, 2016, Sapporo city.

8.

48

11 19 2016

9. Thongprachum

Aksara

48

11 20 2016

3 29 3 2

- 73 -

Adenovirus 53/22

- 74 -

2 2

28

23 2011 6

22 2010 CRE

27 2015

1. 28 2016 6

2. 28 2016 9

27

-

3 2

- 75 -

27

a.

b.

c.

d. b-

e. Etest

f. CarbaNP test g. JANIS h.

i. Clostridium difficile j.

a. Miseq

b. S1-PFGE

c. 1 2

d.

e. S1-PFGE

3.

1.

81 80

a.

1

80%

57

6 47 /80 59%

b.

2

CRE

- 76 -

VRE

- ESBL

B

c.

38

22 12

d.

, ,

-

1 6

3-4

MALDI-TOF MS 1

8

1 MIC Etest 3.5

-

ESBL - - AmpC

-

PCR -

53(66%) 50(63%)

16S rRNA 26(33%)

PCR 30(38%)

58(73%) 3(4%) 8(10%)

Etest 28(35%)

-

ESBL 44(55%)

- - 44(55%)

AmpC - 40(50%)

KPC 23(29%)

CarbaNP 11(14%)

2

- 77 -

IMP NDM VIM KPC OXA-48 6

ESBL 6

OXA-51 OXA-23

4

e.

6-7

58 73

2. 28

28 2016

20 36

27

( 2 )

30

b-

89 24

/27

52% 14 /27

41% 11 /27

33% 9 /27

30 29

24 83%

8 28

10 34

6 21%

3.

25 2013 60

1

21 27 2015

- 78 -

27 2015 28 2016

64 27

2015 17

28 2016 9

8 8

6

23 2011 6

28 2016

6

1500

CRE 100

VRE CRE

VRE 10

CRE

MRSA

CRE

22

-

KPC

28

28

- 79 -

1

1. .

. 28

. 1 20-22 , 2017 , .

2.

.

. 28

. 1 20-22 , 2017 , .

- 80 -