季節性流感防治工作手冊
72
ۑࢬ܄གݯπբЋн ۑࢬ܄གݯπբЋн ۑࢬ܄གݯπբЋн ۑࢬ܄གݯπբЋн Practical Guideline for Prevention and Control of Seasonal Influenza
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有關防治季節性流感相關資訊
Transcript of 季節性流感防治工作手冊
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Practical Guideline for Prevention and
Control of Seasonal Influenza
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2
紺紺紺紺
紺
紺紺紺紺紺紺紺紺紺 18
20 紺
(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紺紺紺紺紺紺紺紺紺紺紺紺紺紺
7 紺
11
22 紺
M2 protein Neuraminidase 40
紺
紺紺紺紺 紺
( ) 56
( ) 57
58 紺
59 紺
60
61 紺
66
70
紺
3
紺紺紺紺
紺紺紺紺� (Seasonal Influenza or Seasonal Flu)
H1N1 H3N2 A / B紺
� 噁噁噁噁Influenza Season or Flu Season)))) 7 1 6 30
� 噁噁噁噁Influenza Epidemic Period)))) 11 3
6 9
� (Influenza-like Illness, ILI) � ( 38 )
� � � (Complicated Influenza )
() ( )
� (Avian Influenza) H5N1 � H5N1 H5N1
� (Pneumonia and Influenza, P&I) 1975 (ICD-9) 480-487
(480) (481) (482)(485) (486) (487)
� (Cluster) 紺紺紺
紺
4
紺紺紺紺
2008
2009 H1N1
2011 9
2011 12
H5N1
IC
Rapiacta
5
(Description of Disease)
(Influenza )
(Influenza virus) 1 4 (1,2)
(1-3) 10
3~7
2 (2,3)
common cold
(Influenza)
(Common Cold)
200
/
3-4
1-2 2-5
( )
( )
6
(Reye syndrome) (1-5)
1933 · Wilson Smith (1-3)
A (H1N1)
A B C A B
A
B
1977 A (H1N1 H3N2)
B
噁Orthomyxoviridae)
A B C Segmented RNA
80-120nm envelope 2
- Hemagglutinin HA, H
Neuraminidase NA, N (1-5) M1
M2 A B 8
(-)RNA (-)RNA Nucleoprotein NP
Transcriptase RNA PB1 PB2 PA C
7 (1,3,4)
(HA NA) A
(H ) (N )
7
H 16 H1-H16 N 9 N1-N9(1-
4) HA NA H1N1 A
HA-1 NA-1 H3N2 A HA-3 NA-
2 B C (1-4)
A B
A H1N1 H2N2 H3N2
H1N1
H7N7 H3N8 B
B C
(1,3,4)
A
1957 1968
( A/H2N2 A/H3N2)
A B C 8 8 7
11 11 9 (Antigenic
drift) (Antigenic shift)
(Antigenic drift) (Antigenic drift)
A
””
8
2009 H1N1
2011 7
H3N2(H3N2 variant) H1N1 M1
H5N1 A/H5N1 H5N1
A 1961 ( )
1997 H5N1
18 6 30 2004
H5N1 59%(5,6) H5N1
H5N1
1. A B 8 RNA C
7
2.
3.
4.
5. Zanamivir(Relenza ) Oseltamivir(Tamiflu )
6.
Reassortment
A B C 1980 (WHO)
H N
9
(1,6) A ⁄ ⁄
⁄ ⁄ (HnNn)
A/swine/Lowa/15/33 (H1N1)
A 1933 Lowa H1N1
15
A/Beijing/262/95 (H1N1) A 1995 262
Beijing H1 HA N1 NA (1-4)
B / / /
HA NA B/Beijing/243/97 C
(1-4)
A
-
(1,3,4)
RNA RNA
" " " "
HA NA
" Antigenic drift " " "
A B
" Antigenic shift " "
"
20-30 (1,3,4) A
48 60
10
5 10
(1,3,6)
(NA)
Shedding
Virema
(1-4)
Mode of transmission 紺紺紺紺
Incubation period
1~4 2
12 3
Period of communicability
3~5 7-10 (1,2)
24-48
24-72 5 (4,6)
(1,2,5)
21
11
Susceptibility and resistance
Antigenic drift (2,5)
(2,5)
A B
A C
Pandemics
(Seasonal epidemics) (Sporadic cases) 20~21
(WHO) 3 5
25 50 (1)
(Centers for Disease Control and Prevention,
CDC) 10 3
Spanish Flu
( )
1918-1919 A/H1N1 4
20-30
Asian Flu
( )
1956-1957 A/H2N2 100 150
Hong Kong Flu
( )
1968-1969 A/H3N2 75 100
Pandemic H1N1
Influenza
(H1N1
)
2009-2010 Pandemic H1N1 1 8 449 (7)
12
10 4
2~4 5
100 (100/100,000)
500/100,000(2,6,8) 2011-2012
3 (9,10)
11 3
4 (EuroFlu) 2010-2011
(Severe acute respiratory infection, SARI)
1,548 H1N1 2011-2012 ( 2012 5
11 ) 1,248 A/H3N2 (9) 紺
Centre for
Health Protection
12
7 8 2011-2012 A/H3N2 (11)
2011-2012 11 2012 1
A/H3N2
(National Notifiable
Diseases Surveillance System(NNDSS)) 5-6
8-9 2011 (1 1 10 16 )
25,092 H1N1
A/H3N2 8 (12)
7 1 6 30
11 12 3 11%
(14)
B A/H3N2 2009 H1N1
13
2009 7 H1N1
2010-2011 A/H3N2 12
H1N1 2011-2012 B A/H3N2
2008-2012
2008-2009 28 2009-2010 1,312
2010-2011 1,785 2011-2012 1,704 7
49 152 H1N1
2008-2009
65 1-6 2009-2010 H1N1
18 0-6 65 (15) 2010-2011
65 0-3 H1N1
0-6 65
14
(http://www.cdc.gov.tw) / /
(http://flu.cdc.gov.tw)
7 10
10-20
40-50
B A
B A Antigenic
drift Antigenic shift
15
1-2
6 65 65
1996 2011 7.7% 10.9%
16
(Surveillance System)
1.
2.
3.
4.
5.
6.
IC
1999
2000
2004
2007
2011 9
17
3 39
( )
* (
) ( )
( )
1.
( )
2. -
(https://ida4.cdc.gov.tw/hospital/)
3.
- (https://ida4.cdc.gov.tw/phb/)
4.
*
1 38
2
3
18
19
( )
( ) ( )
1. Possible case
2. Probable case
3. Confirmed case
(1)
(2) 14 ( )
(3)
( )
(http://nidss.cdc.gov.tw) /
1. ( )
2.
3.
4. 4
20
臨床醫師發現疑似流感併發症個案
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
臨床表現資料是否符合通報條
件
否 不接受通報
是
綜合檢驗結果 可能病例
「附加資訊-
個案狀況維護」是否死亡
極可能病例
【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
「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
否
是
發病日14天(含)內是否曾入住
加護病房治療
確定病例-死亡
否 確定病例-不需加護病房治療否
是** 是
一週內通報
陽性
確定病例-需加護病房治療
*
* : 紺
14 噁 )
紺
**
21
( )
1999 3
255
1.
2.
3.
( )
1.
(1) 38
(2) 3
2.
(1)
(2) 255
3.
2 2012 8
22
4.
(1)
(B)
( )
1.
( )
2.
(A)” ”
Transtube
23
(2)
a. 4
b. 24
c.
( )
(3)
( )
1.
(1)
(2)
a. 4
b. 24
(3) (real-time RT-
PCR)
2.
( )
24
(http://cdc.gov.tw) /
/ / /
( )
紺 紺
IC
Real-time Outbreak and Disease
Surveillance RODS 1999 9
2004 RODS
2006 11
ICD-9-CM
170 10 80%
ICD-9-CM
RODS
2008
IC
IC
RODS
RODS
(http://cdc.gov.tw) / / /
25
(RODS) /RODS
(http://nidss.cdc.gov.tw)/
H1N1 H1N1
2009 4
6
( )
(1) 38
(2)
26
(3)
( )
1.
https://ida4.cdc.gov.tw/hospital/
2.
3.
( )
1.
2.
3.
bar-code (
)
( )
(http://cdc.gov.tw)
/ /
27
https://203.65.72.161/ida2/
1. +
饋.A + H1 H3
2002
665
( )
紺紺紺紺
紺
28
1. ≥ 38°C
2.
( )
1.
2.
3.
4.
5.
6.
( )
(http://cdc.gov.tw) / / /
29
疑似法定傳染病或特殊傳染病
通知校護
通知當地衛生局
衛生局進行調查及後續處理
符合定點學校傳染病通報項目與定義
(註1)
是
通知校護
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、教育
局、疾管局分局
衛生局進行調查及後續處理
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
校護於每下週一下班前上傳該週學生傳染病資料
結案
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 學生生病
疑似法定傳染病或特殊傳染病
群聚事件
是
是 否
通知校護
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、教育
局、疾管局分局
衛生局進行調查及後續處理
否
是
結案
否
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」
否
是
群聚事件
是
結案
否
否
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。
紺紺紺紺
30
Long term care facility The elderly
apartment Institution for disability
Protectory Veteran’s home Prison
Nursing home Day-care center for mental recovery
( )
1.
(1) ≥ 38
(2)
2.
紺
1.
38
37.5
2. 3
( )
1.
(http://iss.cdc.gov.tw)
31
2. 24
3.
10
4.
( )
( )
( )
32
酶
蕁 蕁噁 )
蕁 蕁 ; 蕁 蕁.
(http://www.cdc.gov.tw)/ /
紺紺紺紺
33
( )
1. Nasal swab
2. Nasopharyngeal swab
3. Nasal wash fluid
4. Throat swab
5.
Transtracheal aspirateBronchoalveolar lavage fluidSputum Lung
biopsy Post-mortem lung or Tracheal tissue
6.
Acute serum
Convalescent serum
( )
3 (Nasopharyngeal
aspirate) Throat swab
Immunofluorescence staining
assay, IFA 4
4
48-72
-70
34
( )
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) Real–time RT-PCR
35
RT-PCR Real–time RT-PCR
(Primer)
(RT-PCR ) (Real–time RT-PCR )
( )
Paired sera
10
4
Neutralization test, NT
Hemagglutination inhibition test, HI NT HI Subtype
Strain HI
Immunofluorescence staining assay, IFA
IFA
A B
A H1 H3 H5
Anti-mouse IgG FITC conjugate
Virus Culture
MDCK cell
(HI)
( )
36
RT-PCR Real-time RT-PCR
RT-PCR
RNA PCR
Reverse transcriptase, RT RNA cDNA PCR
H5N1 HA NA
WHO H5 WHO
RT-PCR
RT-PCR
RT-PCR
Internal control
37
2 7
X
10
(2,3,4)
A
B A B
B A
A/H7 A B
(16-18)
(Viral pneumonia)
(Bacterial pneumonia) (Encephalitis) (Encephalophy)
(Myocarditis) (Pericarditis) (Tansverse myelitis)
- (Guillain-Barré syndrome) Reye s syndrome (2-
5)
65
( )
72
38
48
48
65
M2 protein
(Neuraminidase) M2 protein A
neuraminidase A B
( ) M2 protein
Amantadine Rimantadine M2 protein
(ion channel) M2 protein
Amantadine Rimantadine
M2 protein M2 protein
Amantadine
Rimantadine Amantadine
Amantadine Catecholamime Rimantadine
Amantadine 10%
2%
Rimantadine
39
A (
)
( ) (Neuraminidase)
Zanamivir (Relenza TM ) Oseltamivir (Tamiflu® )
Peramivir (Rapiacta®) 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% ) 6-10
99% Rapiacta®
Zanamivir Oseltamivir Peramivir
36-48 1-2
0.4% neuraminidase
4%
噁 )紺M2 protein
40
A Amantadine Rimantadine
( ) Neuraminidase
Oseltamivir Zanamivir
1-1.5
( ) M2 protein
M2 protein
Amantadine Rimantadine
H3N2 H1N1 100%
( ) Neuraminidase
1.5% Oseltamivir
M2 protein Neuraminidase M2
protein A B
Neuraminidase Oseltamivir Zanamivir
A B
M2 protein Neuraminidase
M2 protein Neuraminidase Amantadine Rimantadine Oseltamivir Zanamivir Rapiacta
Symmetrel, Symadine
Flumadine Tamiflu Relenza Peramivir
A A A B A B A B
1 1 1-1.5 1-1.5 1 (10%) (2%) (2%) (1.5%) (90-100%) (90-100%) (1.5%)
41
( ) Oseltamivir (Tamiflu )
13( ) 40
1( )
1. (10 ) 2 5
2. (10 ) 1 10
( ) Zanamivir (Relenza )
5( )
1. 2 (2×5 ) 2 5
2. 2 (2×5 ) 1 10
42
- ACIP
6 18 5 50
body-mass index≥40) <5
≥50 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
43
A H1N1 A H3N2
A B
1
1 T
2
65 70-90%(3-5)
30 70% (Pneumonia and
Influenza, P&I) 30 40%
50~60% 80% (2,5,6)
7 6 12
8 (19)
universal influenza vaccine
12 3 9
11
44
65 6
6
1-6
6 3
0.25 ml 8
1
1 2
Guillain-Barré
1976
Guillain-Barré
45
1.
2. 6
3.
4.
5.
2-8 C
46
1.
2.
3.
1.
IC
2.
1
A.
B.
C.
D.
47
2
(
3
LED
4 (http://www.cdc.gov.tw)
3.
12 3 9
11
2 3
10 1
(http://flu.cdc.gov.tw)/
4.
H5N1
(BMI 35)
/
5.
1 -
2
6.
48
1.
2.
3.
4.
/ 1.
2.
3.
4.
1.
2.
1.
2.
3.
1.
2.
3.
4.
1.
2.
3.
4.
49
12 3
1.
2.
3.
4.
1.
2.
1
A.
B.
C.
D.
2
A.
B.
3
A.
B.
50
3.
48 / /
4.
1
紺
/ 紺 紺
(
)
1.
2.
3.
/
4.
紺
(
)
1.
紺
2. 紺
3. 紺
4. 紺
紺
(
)
1. /
紺
2. 紺
3. 紺
4. 紺
(
)紺
1.
紺
2. 紺
3.
51
( ) 4. 紺
( )
( )
1.
紺
2. 紺
3. 紺
紺
2
a.
( ( )
72 )
b.
48
48
5. 紺
(1) 38
(2)
52
(3) (
)
2
( )
/
( ) ( )
1.
2.
( )
1.
2. /
( ) 48
1. (
)
2.
3. / /
53
( )
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/ ) 紺
54
紺紺紺紺
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
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12. Australian Government Department of Health and Ageing. Australian Influenza Surveillance Report; 2011 No.15
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14. 2002 2004; 20:478-487
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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;29:1844-49.
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Fever 紺紺紺紺 C Headache Myalgia Malaise Rhinorrhea Cough Sore throat Vomiting
Diarrhea Pharyngeal vesicles or ulcers Paralysis Skin rash Others紺 紺 紺 紺 紺 紺 紺 紺紺 紺 紺 紺 紺 紺 紺 紺紺 紺 紺 紺 紺 紺 紺 紺紺 紺 紺 紺 紺 紺 紺 紺
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1 38 2
Bar-code
Throat swab Nasopharyngeal secretion or swab Others____________
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Throat swab Nasopharyngeal secretion or swab Others____________
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______ ______ _______
編號：VL-001 紺紺紺紺
※本項防疫檢驗，不得用以申請健保給付 紺紺 紺
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02-23123456*66908
02-87923311*13568
03-2118800 5497
04-23592525 5701
04-7238595 5930
06-2353535 2653
07-3422121*8098
03-85618255 2090
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1.
� MK-2 cell H292 cell � 200 µl 1 mL
0.45m MDCK 7-10 CPE 3,000rpm15 1 mL PBS 21Acetone Influenza A Influenza B monoclonal antibody
indirect immunofluorescence staining assayIFAapple green
2. Respiratory viruses follow DAKO system – direct IFA � � 10 µl� wet chamber 37 15 � PBS PBS
5 �
*MK-2 cell
MDCK cell
Vero cell
RD cell
HEL cell Hep-2 cell
CPE紺
(+)
(-)
Blind passage
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3. Influenza A virus HA typing by RT-PCR method
RNA
Qiagen Viral RNA Mini Kit 140 µl
50 µl AVE buffer RNA RT
RT follow Promega RT procedure (a). 15 µl RNA 70 5 RNA (b). 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
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 94 2 94 1
40cycles
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52 2 72 3 72 7 4
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 reaction48℃ 30 FluA-F primer 10 µM 1.0 µL Taq activation95℃ 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 0.3 µL Real-time RT-PCR FluA-F primer 10 µM 1.0 µL RT reaction63℃ 3 紺FluA-R primer 10 µM 1.0 µL Taq activation95℃ 30 FluB-F primer 10 µM 1.0 µL PCR reaction FluB-R primer 10 µM 1.0 µL 95℃ 10 FluA probe 5 µM 0.5 µL 58℃ 30 紺FluB probe 5 µM 0.5 µL 72 3 45 cycles Enzyme master mix 7.4 µL Enhancer 1.0 µL Activator 1.3 µL RNA sample 5.0 µL
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紺紺紺紺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 5 destain 5
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