Background

DF/DHF high morbidity and mortality rate Endemic in at least 100 countries A major problem in the world, especialy in tropical countries. Vector control is important, but is reality living habitat of Aedes aegypti is difficult to control Late diagnosis is fatal Mechanism factorr lead to severe dengue is unknown

 Over 50 million infections are reported annually with 250.000-500.000 DHF. Caused by 4 serotype of dengue viruses (Den 1, Den 2, Den 3, Den 4) No antiviral available No vaccine available
WHO. Geneva 2003 Kumarasamy et al. Singapore Med J 2007;48:669-73 Alcon et al. J Clin Microbiol 2002; 40:378-81 Lindergend et al. J.Microbiol 2005;43:2850-55 Lolekha R et al.J Trop. Med. Public Health 2004;35:391-5

NON STRUCTURAL PROTEIN

FUNCTION

NS1 NS2A

Replication Unknown

NS2B NS3

Cofactor NS3, connection on 3 NCR, NS3 and NS5

Protease Serin and NTPase helikase

NS4A NS4B NS5

Replication Unknown Unknown Replication RNA with NS3

STRUCTURAL PROTEIN

FUNCTION

Core Pre M

Nucleocapsid protein Glycoprotein precussor Host cell fussion Assembling of viruses

M protein E Protein

Glycoprotein membrane Host cell fussion of virus Glycoprotein envelope Respon immune induction Antibody reseptor connection

Sophie A et al. J Virol 2005;79:11403-11.

Serotype Den 1 Den 2 Den 3 Den 4

Number of sera 93 31 24 29

Sensitivity of dengue NS1 Ag (95% CI) 88,9%(65,8%-94,8%) 87,1%(70,1%-96,3%) 100%(85,6%-100%) 93,3%(77,9%-97,9%)

Philippe D et al. Clin and Vaccine 2006; 13:1185-89 Alcon S et al. J Clin. Microbiol 2002; 40:378-81 Chou-Feng et al.J Virol 2005;79:11403-11

Serotype Den 1 Den 2

No.of sera test 42 42

No(+) NS1 Ag test 38 37

Sensitivity (%) 90,5 88.1

95% CI 77,4-97.3 77.4-96

Den 3
Den 4 All serotype

107
48

94
43

87.9
89.6

80.1-93.4
77.3-96.5

239

212

88.7

84-92.4

Chung Yi L et al. Am J Trop Med Hyg 2003;32:164-69 Paul R et al. J Clin Microbiol 2000;38:1053-57.

Day after begining of fever

0

Number of sera

Sensitivity of Dengue NS1 antigen (%)

100

Sensitivity of Dengue IgM ELISA(%)

0

10

1
2 3 4

33
40 20 27

87.8
92.5 95 96.3

5.1
6.1 15 48.1

5
6

19
28

52.6
35.7

94.1
100

Paul R et al. J Clin Microbiol 2000;38:1053-57 Pei Yun S et al. Clin and Diags Laboratory Immunol 2003;10:622-30

Diagnostic methode

Number of serum samples tested positive

Primary Dengue (NS1) Secondary Dengue (NS1) 7(6) 20(12) 2(2) 3(2) Combined (NS1) 71(68) 68(57) 74(74) 11(10)

Viral isolation alone RT-PCR Viral isolation + RT-PCR Serology (pair sera)

64(62) 48(45) 72(72) 8(8)

Total

192(187)

32(22)

224(209)

Alcon S et al. J Clin Microbiol 2002;40:378-81 Shu P et al. Clin Diags Laboratory Immunol 2004;11:642-50. Pei Yun S et al.Clin Diags and Laboratory Immunol 2003;10:622-30.

Diagnostic methode

Number of serum samples tested positive

Primary dengue (NS1) Secondary dengue (NS1) Combined (NS1)

Viral isolation alone RT PCR Viral isolation+RT-PCR Serology(pair sera) Total

29(28) 13(12) 18(18) 8(8) 68(66)

5(4) 12(6) 0(0) 3(2) 20(12)

34(32) 25(18) 18(18) 11(10) 88(78)

Diagnostic methode

Number of serum samples tested positive

Primary dengue (NS1) Secondary dengue (NS1) Combined (NS1)

Viral isolation alone RT PCR Viral isolation+RT-PCR Serology(pair sera) Total

35(34) 35(33) 54(54) 0(0) 124(121)

2(2) 8(6) 2(2) 0(0) 12(10)

37(36) 43(39) 56(56) 0(0) 136(131)

Eric M et al. Am J Trop Med Hyg 2003;32:164-69 Darfu et al. Clin Chemistry 2006;52:1486-91 Lindergend G et al 2005; 43:2850-5

Day of fever

No. Sera of tested

Dengue NS1 Antigen Test No. (+) test 15 61 54 34 19 8 Sensitvity (%) 100 87.1 91.5 85 76 42.1 95% CI 78.2-100 77-93.9 81.3-97.2 70.2-94.3 54.9-90.6 20.3-66.5 No. (+) Elisa 0 1 3 6 8 16

MAC ELISA Sensitivity (%) 0 1.4 5.1 15 32 84.2 95% CI 0.0-23.2 0-7.6 1.1-14.2 5.7-29.8 15-53.5 60.4-96.6

0 1 2 3 4 5

15 70 59 40 25 19

6
7 Total

18
20 266

17
9 217

94.4
45 81.6

75.7-99.8
23.1-68.5 76.4-86.1

18
20 72

100
100 26.7

81-100
83.1-100 21.8-32.8

Kumarasamy V et al. Singapore Med J 2007;48:669-73 Pei Yun S et al. Clin Diags and laboratory Immunol 2003;10:622-30 Pei YS et al. J Clin Microbiol 2002; 40:1840-44.

Dengue is endemic desease affecting tropical and subtropical region. Infections with dengue virus causes a broad spectrum of illnesses. Early detection and management is important Prompt diagnosis of dengue infection is imperative in the control of dengue virus infection and management of patient. Detection nonstructural protein NS1 antigen an alternative methode for early diagnosis. Sensitivity 88%-100% and spesificity 100%.