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紺 紺 Practical Guideline for Prevention and Control of Seasonal Influenza 紺

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4 5 5 6 紺紺紺

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1


紺 紺 紺紺紺紺紺紺紺紺紺

18 紺

20 (A)

(B)

22 27 29 32 紺 5

紺紺紺紺紺紺紺紺紺紺紺紺紺紺紺紺紺紺紺紺

7

11 紺

22 M2 protein

Neuraminidase

40

紺 紺 (

) (

56 )

57 58

59

60 61 66 70 紺

2


紺 紺 

(Seasonal Influenza or Seasonal Flu) H1N1

H3N2

A

/

B

紺 噁Influenza Season or Flu Season) )

 7

1

6

30

噁Influenza Epidemic Period) )



11 6



9

(Influenza-like Illness, ILI) 

(







(Complicated Influenza ) ( )

(



)

(Avian Influenza) H5N1

 H5N1 H5N1



(Pneumonia and Influenza, P&I) 1975 480-487

(ICD-9) (480) (485)



3

(486)

(481) (487)

(482)

(Cluster)

紺紺紺 紺

3

38

)


ç´ş

2008

2009

H1N1 2011

2011

9

12

H5N1

IC

Rapiacta

4


(Description of Disease)

(Influenza ) (Influenza virus)

(1-3)

1

4

(1,2)

10 3~7 (2,3)

2

common cold

(Influenza)

(Common Cold) 200

/ 3-4 1-2

2-5

(

)

5

(

)


(1-5)

(Reye syndrome)

1933

·

(1-3)

Wilson Smith

A

(H1N1)

A B C

A

B

A B 1977

A

(H1N1

H3N2)

B

噁Orthomyxoviridae) A

B

C

Segmented

RNA

80-120nm -

Hemagglutinin

Neuraminidase

NA,

M2

B

A

HA,

2

H

(1-5)

N

M1 8

(-)RNA Transcriptase 7

envelope

(-)RNA

RNA

Nucleoprotein

PB1

PB2

PA

NP

C

(1,3,4)

(HA (H

)

(N

6

)

NA)

A


H 4)

16

HA

HA-1

NA

B

N

H3N2

N1-N9(1-

9

H1N1

NA-1

2

H1-H16

A

A

HA-3

NA-

(1-4)

C A

B

A

H1N1

H2N2

H3N2 H1N1

H7N7

H3N8

B

B C

(1,3,4)

A

B

8 11

C

8 11

7 9

(Antigenic (Antigenic drift)

drift) (Antigenic shift)

A

” ”

A 1957 (

1968

A/H2N2

A/H3N2)

7

(Antigenic drift)


2009

H1N1 2011

H3N2(H3N2 variant)

H5N1

H1N1

A/H5N1 A

M1

H5N1

1961

( 1997

18

7

6

)

H5N1

30

2004 59%(5,6)

H5N1

H5N1

H5N1

1.

A

B

8

RNA

C

7 2.

3.

4.

5.

Zanamivir(Relenza

)

Oseltamivir(Tamiflu

)

6. Reassortment

A B C H

1980 N

8

(WHO)


(1,6)

A

(HnNn) A/swine/Lowa/15/33 (H1N1)

A

1933

Lowa

H1N1

15 A/Beijing/262/95 (H1N1)

A

Beijing

H1

B

HA

/ HA

1995 N1

NA

(1-4)

NA

/

262

/

B/Beijing/243/97

C

(1-4)

A -

(1,3,4)

RNA

RNA "

HA

"

"

NA

" A

"

Antigenic drift

"

"

"

B

"

Antigenic shift

" (1,3,4)

20-30

48

60

9

A

"

"


5

10

(1,3,6)

(NA) Shedding

Virema (1-4)

ç´ş

Mode of transmission

Incubation period 1~4

2

12

3

Period of communicability 3~5

7-10

24-48 24-72

5 (1,2,5)

21

10

(4,6)

(1,2)


Susceptibility and resistance

(2,5)

Antigenic drift

(2,5)

A

B A

C

Pandemics (Seasonal epidemics)

Spanish Flu (

1918-1919

(Sporadic cases)

A/H1N1

)

Asian Flu

4 20-30

1956-1957

A/H2N2

100

Hong Kong Flu 1968-1969

A/H3N2

75

(

20~21

150

)

(

100

)

Pandemic H1N1 2009-2010

Pandemic H1N1

1

8

(7)

449

Influenza (H1N1 ) (WHO) 25

50

3

5

(1)

(Centers for Disease Control and Prevention, 10

CDC)

11

3


10

4 2~4

100

5

(100/100,000) 500/100,000(2,6,8) 2011-2012 (9,10)

3 11 4

(EuroFlu)

3

2010-2011

(Severe acute respiratory infection, SARI) 1,548

H1N1

11

)

2011-2012

(

2012

5

A/H3N2 (9) ç´ş

1,248

Centre for Health Protection 12 7

8

2011-2012

2011-2012

(11)

A/H3N2 11

2012

1

A/H3N2 (National Notifiable Diseases Surveillance System(NNDSS)) 8-9

2011

(1

1

5-6 10

16

25,092

H1N1

A/H3N2

(12)

8

7 11

)

12

1

6

30

3

11%

(14)

B

A/H3N2

12

2009

H1N1


2009 2010-2011

7

H1N1

A/H3N2

H1N1

12

2011-2012

B

A/H3N2

2008-2012

2008-2009

28

2010-2011 49

1,785

2011-2012

2009-2010

1,312

1,704

152

7

H1N1 2008-2009

65

1-6 18

65

0-6 0-3

2009-2010

H1N1

65

(15)

H1N1 0-6

13

65

2010-2011


(http://www.cdc.gov.tw)

/

/

(http://flu.cdc.gov.tw)

7

10

10-20 40-50

B B drift

A A

Antigenic shift

14

Antigenic


1-2

6 1996

65 2011

65 7.7%

10.9%

15


(Surveillance System)

1.

2. 3. 4. 5. 6.

IC

1999 2000 2004 2007 2011

9

16


3

(

39

)

*

( )

(

)

* 1

38

2 3 (

) 1.

( ) 2.

(https://ida4.cdc.gov.tw/hospital/)

3. -

(https://ida4.cdc.gov.tw/phb/)

4.

17


18


(

)

(

1.

)

(

)

Possible case

2.

Probable case

3.

Confirmed case (1)

(2)

14

(

(3)

1.

(

)

2. 3. 4.

(

4

)

(http://nidss.cdc.gov.tw) /

19

)


臨床醫師發現疑似流感併發症個案 一週內通報

衛生局接獲通報個案

【衛生局、醫院】 1. 收集檢體(咽喉擦拭液) 2. 進行檢體送驗事宜

【衛生局】 1. 檢核醫師以網路或書面(須協助登錄通 報系統)通報之資料,包括臨床表現、 流感疫苗接種史及流行病學接觸史等 2. 定期追蹤及維護個案狀況(入住加護病 房及死亡情形),以確實完成資料

檢體請依標準作業程序運送

【本署認可實驗室或本局呼吸道病毒實驗室】 1. 進行相關檢體檢驗事宜 2. 上網登錄檢驗結果

【傳染病個案通報系統】 依據附加資訊中之「臨床表現」、「流行病學 」資料及「綜合檢驗結果」與「入住加護病房 及死亡情形」進行個案自動研判

臨床表現資料 是否符合通報條 件

不接受通報

綜合檢驗結果

「附加資訊 -流行病學接觸史」 是否有流行病學 接觸史

陰性或 超過21日 無檢驗結果

陽性

可能病例

極可能病例

* 「附加資訊個案狀況維護」是 否死亡

*

:

發病日14天 (含)內是否曾入住 加護病房治療

是**

確定病例-死亡

確定病例-需加護病房治療

確定病例-不需加護病房治療

14

) 紺

**

20


(

) 1999

3

255

1.

2.

3. (

) 1.

(1)

38

(2)

3

2. (1) (2)

255

3.

2

2012

21

8


4. (1)

1. (

) (

)

2.

(A) ” ”

Transtube

(B)

22


(2) a.

4

b.

24

c. )

(

(3) (

) 1. (1) (2) a.

4

b.

24

(3)

(real-time RTPCR)

2.

(

)

23


(http://cdc.gov.tw) /

(

/

/

/

) 紺

IC Real-time Outbreak and Disease Surveillance RODS

1999

9

2004 2006

RODS

11

ICD-9-CM

170

10

80%

ICD-9-CM

RODS 2008 IC IC

RODS

RODS (http://cdc.gov.tw)

/

/

/

24


(RODS)

/RODS (http://nidss.cdc.gov.tw)/

H1N1

H1N1 2009

4

6

(

)

(1)

38

(2)

25


(3) (

) 1.

https://ida4.cdc.gov.tw/hospital/ 2.

3.

(

) 1. 2.

3. bar-code

( )

(

) (http://cdc.gov.tw) /

/

26


https://203.65.72.161/ida2/

1. 饋.A

2002 665

(

)

27

+ +

H1

H3


1.

≥ 38°C

2.

(

) 1.

2.

3.

4.

5.

6. (

)

(http://cdc.gov.tw)

/

28

/

/


紺 各級公私立學校

是否為「定點傳染病監視 通報系統」之定點學校

學生生病

疑似法定傳染病 或特殊傳染病

學生生病

結案

校護於每下週一 下班前上傳該週 學生傳染病資料

群聚事件

否 通知校護

群聚事件

通知校護

符合定點學校傳染 病通報項目與定義 (註1)

導師每日登記 「學生感染疑似 傳染病登記週報 表」,於每週五 下班前交給校護

疑似法定傳染病 或特殊傳染病

通知校護

若為群聚事件則 需 24小時內電話或 填寫「學校疑似 傳染病群聚速報 單」

結案 24小時內電話 或填寫「學校 疑似傳染病群 聚速報單」

是 24小時內電話 或填寫「學校 疑似傳染病群 聚速報單」

通知當地 衛生局

通知當地 衛生局、教育 局、疾管局分局

衛生局進 行調查及 後續處理

結案

通知當地 衛生局、教育 局、疾管局分局

衛生局進行調 查及後續處理

衛生局進行調 查及後續處理

註1:定點學校傳染病通報項目與定義 *類流感:急性呼吸道感染且具有下列症狀:1.突然發病有發燒(耳溫380C)及呼吸道感染2.且有肌肉酸痛或頭 痛或極度厭倦感。 *手足口病或疱疹性咽峽炎:手足口病:口、手掌、腳掌及或膝蓋、臀部出現小水泡或紅疹;疱疹性咽峽炎:發 燒、咽部出現小水泡或潰瘍。 *腹瀉:每日腹瀉三次(含)以上,且合併下列任何一項以上症狀者:1.嘔吐 2.發燒3.黏液狀或血絲 4.水瀉。 *發燒:耳溫量測超過380C,且未有符合上述「類流感」、「手足口病或疱疹性咽峽炎」、「腹瀉」症狀者。 *紅眼症:眼睛刺痛、灼熱、怕光、易流淚、異物感、霧視;眼結膜呈鮮紅色,有時會有結膜下出血;眼睛產生 大量黏性分泌物;有時耳前淋巴結腫大、壓痛 *其他特殊傳染病,並視疫情狀況調整通報項目。

29


Long term care facility apartment

Institution for disability Protectory

Veteran’s home

Nursing home

(

The elderly

Day-care center for mental recovery

)

1. ≥ 38

(1) (2)

2.

紺 1. 38 37.5 2.

(

Prison

3

) 1. (http://iss.cdc.gov.tw)

30


2.

24

3. 10

4. (

)

(

)

(

)

31


;

)

.

(http://www.cdc.gov.tw)/

32

/


(

) 1.

Nasal swab

2.

Nasopharyngeal swab

3.

Nasal wash fluid

4.

Throat swab

5. Transtracheal aspirate Bronchoalveolar lavage fluid biopsy

Sputum

Lung

Post-mortem lung or Tracheal tissue

6. Acute serum Convalescent serum

(

)

3 aspirate)

(Nasopharyngeal

Throat swab

Immunofluorescence staining assay, IFA

4 4 48-72 -70

33


( ) 1.

10g veal infusion broth

2g bovine albumin fraction V

400 ml 2.

0.8 ml gentamicin sulfate solution (50mg/ml)

3.2 ml amphotericin

B (250ug/ml) 3. ( )

(

)

0.8

NaCl

Rapid antigen detection Enzyme immunoassay Nucleoprotien, NP

A

B

15-30

(

)

Virus culture 3-10

MDCK

(Madin-

Darby Canine Kidney cell) Cytopathic Effect, CPE

Hemagglutination- inhibition, HI

(

)

RT-PCR

Real-time RT-PCR

3~10 Reverse transcriptase-Polymerase chain reaction (RT-PCR)

34

Real–time RT-PCR


RT-PCR

Real–time RT-PCR

(Primer) (RT-PCR

(

)

(Real–time RT-PCR

)

)

Paired sera 10 4

Neutralization test, NT Hemagglutination inhibition test, HI Strain

NT HI

Subtype

HI

Immunofluorescence staining assay, IFA IFA A A

H1

H3

B H5

Anti-mouse IgG FITC conjugate

Virus Culture

MDCK cell (HI) (

)

35


RT-PCR

Real-time RT-PCR

RT-PCR RNA Reverse transcriptase, RT

PCR RNA

cDNA

H5N1

PCR HA

WHO H5

NA

WHO

RT-PCR RT-PCR

RT-PCR Internal control

36


2

7

X

10 (2,3,4)

A B

A B

B

A

A/H7

A

B

(16-18)

(Viral pneumonia) (Bacterial pneumonia) (Myocarditis) -

(Encephalitis)

(Pericarditis)

(Encephalophy) (Tansverse myelitis)

(Guillain-BarrĂŠ syndrome)

Reye

s syndrome

5)

65 ( 72

37

)

(2-


48 48 65

M2 protein (Neuraminidase)

M2 protein

neuraminidase

A

A B

( ) M2 protein Amantadine

Rimantadine

M2 protein

(ion channel)

M2 protein Amantadine

M2 protein

Rimantadine

M2 protein

Amantadine Rimantadine

Amantadine

Amantadine

Catecholamime

Amantadine

Rimantadine 10% 2%

Rimantadine

38


A

(

)

( )

(Neuraminidase) Zanamivir (Relenza TM

Peramivir (Rapiacta®)

) Oseltamivir (Tamiflu®

A

)

(Hemagglutinin)

(Neuraminidase)

N-acetylneuraminic acid (

Sialic acid)

hemagglutinin neuraminidase Sialic acid neuraminidase neuraminidase

A

B

Zanamivir

78% 15%

5-15% 2% Oseltamivir

oseltamivir carboxylate (

75%

) Rapiacta®

99%

Zanamivir

Oseltamivir

Peramivir

36-48 0.4%

1-2

neuraminidase

4%

6-10

)紺M2 protein

39


A

Amantadine

Rimantadine

( ) Neuraminidase Oseltamivir

Zanamivir 1-1.5

(

) M2 protein M2 protein Amantadine H3N2

Rimantadine

H1N1

100%

( ) Neuraminidase 1.5% Oseltamivir

M2 protein protein

Neuraminidase

M2

A

B

Neuraminidase A

Oseltamivir

Zanamivir

B M2 protein

M2 protein Amantadine Rimantadine Symmetrel, Flumadine Symadine A A

1 (10%) (90-100%)

Neuraminidase Neuraminidase Oseltamivir Zanamivir Tamiflu Relenza A

1 (2%) (90-100%)

B

1-1.5 (2%) (1.5%)

40

A

B

1-1.5 (1.5%)

Rapiacta Peramivir A

B

1


( ) Oseltamivir (Tamiflu

)

13( )

40 1( )

1.

(10

)

2

2.

(10

)

1

( ) Zanamivir (Relenza

5 10

)

5( )

1.

2

(2Ă—5

)

2

5

2.

2

(2Ă—5

)

1

10

41


-

ACIP 6

18

5

50

body-mass index≼40)

<5

â&#x2030;Ľ50

6 65 6

Inactivated trivalent influenza vaccines, TIV

(Live-

attenuated influenza vaccine, LAIV)(2,6)

Whole virus vaccine Split-virus vaccine Subunit vaccine Purified surface-antigen vaccine HA

NA (cell based) influenza virus-like particle, VLP WHO

83

110 2

42


A

H1N1

A

H3N2

A

B

1 1

T

2

70-90%(3-5)

65 30

70%

Influenza, P&I)

30

50~60%

(Pneumonia and 40%

(2,5,6)

80%

7

6

12 (19)

8

universal influenza vaccine

12

3

9 11

43


65

6

6

1-6

6

3 0.25 ml

8

1

1

2

Guillain-Barré 1976 Guillain-Barré

44


1. 2.

6

3. 4. 5.

2-8 C

45


1. 2. 3.

1.

IC

2. 1 A.

B.

C.

D.

46


2 ( 3 LED 4

(http://www.cdc.gov.tw)

3. 12

3

9

11

2 10

(http://flu.cdc.gov.tw)/ 4. H5N1

(BMI 35) / 5. 1

-

2

6.

47

3 1


1.

2. 3. 4. /

1. 2.

3. 4.

1. 2.

1. 2. 3. 1. 2. 3. 4. 1. 2. 3. 4.

48


12

1. 2. 3. 4.

1.

2. 1 A. B. C.

D.

2 A. B. 3 A.

B.

49

3


3.

48

/

/

4. 1 紺 /

紺 1.

(

2.

3. )

/ 4.

1.

(

紺 )

2. 紺 3. 紺 4. 紺

紺 /

1.

(

紺 )

2. 紺 3. 紺 紺

4. 1. (

紺 2. )紺

紺 3.

50


( ) ( ) (

4. 1.

)

紺 2. 紺 3. 紺

紺 2

a. (

( 72

) )

b. 48 48

5.

(1)

38

(2)

51


(3)

( )

2

( ) /

( )

(

)

1.

2.

( ) 1. 2.

( )

/

48

1.

( )

2.

3.

/

52

/


( ) 1. (1)

(2) A. B.

C. 2. /

3. (1) (2) 4. 紺紺紺紺紺紺紺 紺紺紺紺 5. 6.

噁http://eir.cdc.gov.tw/ ) 7. (1) 7 (2) 噁http://eir.cdc.gov.tw/ )

53


1. World Health Organization.The world health report 2007 : a safer future : global public health security in the 21st century. World Health Organization; 2007: 45-48. 2. Mandell, GL., Bennett, JE., and Dolin, R. ‘Principles and Practice of Infectious Diseases’, 6th eds. Elsevier Inc. 2005: 2060-2078. 3. U.S. Centers for Disease Control and Prevention, National Center for Prevention Services. ‘Prevention and Control of Vaccine-Preventable Diseases: Influenza’, 7th eds. U.S. Dept. of Health and Human Services. 2003; 2006: 235-56. 4. Cox N.J. and Subbarao K. Influenza. Lancet; 1999;354: 1277-82. 5. Nicholson K.G., Wood J.M. and Zambon M. Influenza. Lancet; 2003; 362:1733-45. 6. U.S. Centers for Disease Control and Prevention. Prevention and control of Influenza.Recommendations of the Advisory Committee on Immunization Practices (ACIP). MMWR 2006;55(RR 10):1-42. 7. World Health Organization. Global Alert and Response (GAR). Pandemic (H1N1) 2009-update 112. Available at: http://www.who.int/csr/don/2010_08_06/en/index.html 8. World Health Organization. Influenza. Available at: http://www.who.int/csr/disease/influenza/en/ 9. World Health Organization. Weekly Epidemiological Record. Review of the 20112012 Winter Influenza Season, Northern Hemisphere; 2012; 87:233-240 10. Centers for Disease Control and Prevention. 2011-2012 Flu Season Draws to a Close. Available at : http://www.cdc.gov/flu/spotlights/2011-2012-flu-seasonwrapup.htm 11. Sentinel Surveillance. Centre for Health Protection in Hong Kong. Monthly summary tables of influenza virus isolation 2011, 2012. Available at http://www.chp.gov.hk/sentinel.asp?lang=tc&id=292&pid=44&ppid=26 12. Australian Government Department of Health and Ageing. Australian Influenza Surveillance Report; 2011 No.15 13. World Health Organization. Cumulative number of confirmed human cases of avian influenza A(H5N1) reported to WHO. Available at : http://www.who.int/influenza/human_animal_interface/H5N1_cumulative_table_ar chives/en/ 14. 2002 2004; 20:478-487 15. 2010 2012; 28:113-126 16. 2005/2006 2007;23:487-502 17. Falsey AR, Hennessey PA, Formica MA, Cox C, Walsh EE: Respiratory syncytial virus infection in elderly and high-risk adults. N Engl J Med 2005; 352: 1749-59. 18. Peltola V, Ziegler T, Ruuskanen O: Influenza A and B virus infections in children. Clin Infect Dis. 2003; 36: 299-305. 19. Masahiko Katayose et al. The effectiveness of trivalent inactivated influenza vaccine in children over six consecutive influenza seasons. Vaccine 2011;29:184449.

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55


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編號:VL-001

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紺 紺 Fever

紺 C

Diarrhea

紺 紺 紺 紺

Headache

Myalgia

Pharyngeal vesicles or ulcers

Malaise

Rhinorrhea

Paralysis

Skin rash

Cough

Sore throat

Vomiting

Others紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 紺 1

38

2

Bar-code

Throat swab Nasopharyngeal secretion or swab Others____________ Throat swab Nasopharyngeal secretion or swab Others____________

/

/

/

/ ______

______ _______

57

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紺紺紺紺紺紺紺紺疱疱.08.饋6

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※本項防疫檢驗,不得用以申請健保給付


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06-2353535

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紺 1. *MK-2 cell

MDCK cell Hep-2 cell HEL cell Vero cell RD cell

(+) CPE紺

 MK-2 cell H292 cell  200 µl 0.45m MDCK 15 Acetone Influenza A Influenza B indirect immunofluorescence staining assay IFA apple green 2.

Respiratory viruses   5 

Blind passage

(-)

1 mL

follow DAKO system – direct IFA 10 µl wet chamber 37

66

7-10 CPE 1 mL PBS monoclonal antibody

15



PBS

3,000rpm 21

PBS


3.

Influenza A virus HA typing by RT-PCR method RNA Qiagen Viral RNA Mini Kit 140 µl 50 µl AVE buffer RT (a). (b).

RNA

follow Promega RT procedure 15 µl RNA 70 40 µl RNA 15 µl 5X buffer 5 µl 100mM dNTP mix 0.5 µl RT 1 µl Random primer 1 µl H2O 17.5 µl

RT 5

RNA

PCR follow Qiagen PCR core kit procedure a. PCR 5X Q solution 10 µl 10X buffer 5 µl 25mM MgCl2 4 µl 10mM dNTP 4 µl RT template 2 µl A/H1 (H3)-1 10 µM 1 µl A/H1 (H3)-2 10 µM 1 µl Taq polymerase 0.5 µl H2O 22.5 µl b. PCR program 40cycles 94 2 94 1

67


52 72 72 4

2 3 7

FLUA/B ABI

(fiollow TaqMan one-step RT-PCR master mix reagents procedure) Real-time RT-PCR 2X Master mix buffer 12.5 µL RT reaction 48℃ 30 FluA-F primer 10 µM 1.0 µL Taq activation 95℃ 10 FluA-R primer 10 µM 1.0 µL PCR reaction FluB-F primer 10 µM 1.0 µL 95℃ 15 FluB-R primer 10 µM 1.0 µL 60℃ 1 45 cycles FluA probe 5 µM 0.5 µL FluB probe 5 µM 0.5 µL RNA enzyme mix 0.67 µL RNase-free water 1.83 µL template 5.0 µL Roche

(LightCycler 480 RNA master hydrolysis probes reagents procedure)

RNase-free water FluA-F primer 10 µM FluA-R primer 10 µM FluB-F primer 10 µM FluB-R primer 10 µM FluA probe 5 µM FluB probe 5 µM Enzyme master mix Enhancer Activator RNA sample

0.3 µL 1.0 µL 1.0 µL 1.0 µL 1.0 µL 0.5 µL 0.5 µL 7.4 µL 1.0 µL 1.3 µL 5.0 µL

Real-time RT-PCR RT reaction 63℃ 3 Taq activation 95℃ 30 PCR reaction 95℃ 10 58℃ 30 紺 72 3 45 cycles

68


紺 Primer A/H1 H1-1: 5’-GAT GCA GAC ACA ATA TGT AGA GG-3’ Primer A/H1 H1-2: 5’-CNC TAC AGA GAC ATA AGC ATT T-3’ Primer A/H3 H3-1: 5’-TCA GAT TGA AGT GAC TAA TGC T-3’ Primer A/H3 H3-2: 5’-AAT TTT GAT GCC TGA AAC CGT-3’ FluA-F :5’- AAG ACC AAT CCT GTC ACC TCT GA -3’ FluA-R :5’- CAA AGC GTC TAC GCT GCA GTC C -3’ FluB-F: 5’- GAG ACA CAA TTG CCT ACC TGC TT -3’ FluB-R: 5’ – TTC TTT CCC ACC GAA CCA AC -3’ FluA probe- FAM -5′ TTT GTG TTC ACG CTC ACC GT 3′-TAMRA FluB probe -VIC-5′AGA AGA TGG AGA AGG CAA AGC AGA ACT AGC 3′-TAMRA FluA probe: 5’FAM - TTTGTGTTCACGCTCACCGT –3’BBQ FluB probe: 5’HEX-AGAAGATGGAGAAGGCAAAGCAGAACTAGC Run gel and read data 1.5 agarose gel 100

40

Ethidium bromide

紺 紺 紺 紺 紺 紺 紺 紺

69

5

destain 5


/ 1

2

( )

3 (1)

38

(2) (3)

70


(

) /

(

) MIS 201

(

)

71


季節性流感防治工作手冊