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Evolution of Enterovirus 71. Jen-Ren Wang, Ph. D. ( 王貞仁 ) Dept. of Medical Laboratory Science and Biotechnology, National Cheng Kung University NHRI Tainan Virology Laboratory for Diagnosis and Research Taiwan CDC Virology Contract Laboratory. Outline. Enterovirus surveillance in Taiwan

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Evolution of Enterovirus 71

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Evolution of enterovirus 71

Evolution of Enterovirus 71

Jen-Ren Wang, Ph. D. (王貞仁)

  • Dept. of Medical Laboratory Science and Biotechnology, National Cheng Kung University

  • NHRI Tainan Virology Laboratory for Diagnosis and Research

  • Taiwan CDC Virology Contract Laboratory


Evolution of enterovirus 71

Outline

  • Enterovirus surveillance in Taiwan

  • Molecular analysis of EV71

  • Virulent Gene Analysis of Enterovirus 71 and Disease Association


Evolution of enterovirus 71

Taiwan CDC and Virology Contract Laboratories

Taipei Veterans General Hospital

Chang Gung Memorial Hospital, Linkou

Tri-Service General Hospital

Taiwan University Hospital

China Medical University Hospital

Buddhist Tzu Chi General Hospital

Chang Gung Memorial Hospital, Kaoshiung

Cheng Kung University Hospital

Kaoshiung Medical University Hospital

CDC, Taiwan

Taichung Veterans General Hospital

Chunghua Christian Hospital

Kaoshiung Veterans General Hospital


Evolution of enterovirus 71

Enterovirus positive cases, 2000-2005 Taiwan

(Virology contract Labs)


Evolution of enterovirus 71

Enteroviruses isolated in contract labs-I

2000 (n=1310)2001 (n=2114)2002 (n=1839)

RankSerotype%Serotype%Serotype%

1Untyped44.4Untyped43.8Untyped45.9

2EV 7118.2CA 1615.5CA 1616.2

3CA 1617.1EV 7115.0E 610.2

4CB 34.9E 3012.1CB 59.4

5E 94.6E 65.4EV 717.9

6CB 44.0P3.8CA 243.3

7E 41.3CB 41.6CB 22.3

8P0.8CA 240.8P1.6

9CA 40.7CA 90.5E 90.8

10E 30.7CB 50.5E 40.5

11CB 10.6CA 100.3CB 40.5

12CA 240.5CA 60.1CA 100.4

13CA 90.3CB 10.1CA 40.3

14CA 20.3CB 30.1E 110.2

15 most frequent

serotypes98.499.699.5


Evolution of enterovirus 71

Enteroviruses isolated in contract labs-II

2003 (n=2066)2004 (n=2389)2005 (n=2031)

RankSerotype%Serotype%Serotype%

1CA 1635.8CA 423.8CB 330.6

2Untyped17.7CA 1022.7CA 1625.5

3EV 7113.0CB 416.5EV 7113.9

4E 97.5Untyped9.8CA 66.6

5E 116.2EV 718.5CA 55.9

6CA 24.7CA 62.6Untyped5.3

7CA 63.9CA 92.6CA 23.8

8CA 53.2CB 32.2E 93.3

9CB 22.0CA 52.0CB 21.4

10CB 51.3CA 21.8CA 101.0

11P0.9CA 161.7P0.9

12CA 110.6CB 21.2CA 40.6

13E 300.6E 251.0CA 90.5

14CA 100.5E 110.9CB 10.2

15 most frequent

serotypes97.997.399.5


Clinical presentations of ev71

Clinical presentations of EV71

  • Infection of EV71 can cause hand-foot-and-mouth disease (HFMD), herpangina, aseptic meningitis, acute flaccid paralysis (AFP), brainstem encephalitis, and pulmonary edema.

  • Several death-associated outbreaks of EV71 infections occurred in Taiwan from 1998 to 2005.


Enterovirus 71 ev71

Enterovirus 71(EV71)

1975, Bulgaria

1978, Hungary

1997, Japan

1969, California

1998, Taiwan

1998, Singapore

1997, Malaysia

1999, Australia

Ho, M, et al. Engl J Med. 1999.


Ev71 in west pacific region

EV71 in West Pacific region

  • Large EV71 epidemics were also reported in Malaysia, Japan, Singapore, Australia, China, Korea, India.


Evolution of enterovirus 71

Isolations of CA16 and EV71


Evolution of enterovirus 71

Outline

  • Enterovirus surveillance in Taiwan

  • Molecular analysis of EV71

  • Virulent Gene Analysis of Enterovirus 71 and Disease Association


Evolution of enterovirus 71

1986

1998

1999

2000

2001

2002

2003

2004

2005

CA16

A

BrCr

7423-MS-87

72

237-TW-86

Phylogenetic analysis of EV71 isolates in Taiwan from 1986 and 1998-2005 based on VP1 region

Both genotypes were able to cause mild and severe diseases.

242-TW-86

99

99

N2838-TW-03

9-97-SHA89

99

5145-TW-98

236-TW-86

97

99

N3793-TW-01

99

N0991-TW-01

S0320-TW-02

81

B

N2443-TW-02

N3496-TW-02

5666-SIN-002209

99

91

5865-SIN-000009

99

N0781-TW-01

N1643-TW-00

H0148-TW-00

80

S0875-TW-99

S0942-TW-99

S0943-TW-99

99

M0167-TW-00

N0716-TW-00

M0201-TW-00

99

S18191-SAR-02

02363-Thai-02

93

1814-JAP-01

1585-JAP-01

99

99

1695-JAP-01

2086-TW-98

99

5746-TW-98

H0001-TW-98

H0358-TW-98

NCKU9822

6092-TW-98

99

H0139-TW-98

4643-TW-98

1245a-TW-98

4-SHZH-01

C

2779-JAP-02

N3340-TW-02

SHZH03

99

40-SHZH-02

03-SHZH-01

08-SHZH-01

99

62-SHZH-02

N2735-TW-04

S0584-TW-04

N2121-TW-05

99

N2472-TW-04

N1862-TW-05

N2490-TW-04

N1614-TW-04

N0003-TW-05

N3031-TW-04

N2411-TW-04

0.1

0.1


Evolution of enterovirus 71

Phylogenetic tree for VP1 region of EV71 strain isolated in Yamagata, Japan, between 1998 and 2003 and reference strains.

Mizuta et al, J Clin Micro 43: 6171, 2005


Evolution of enterovirus 71

Phylogenetic analysis of EV71 based on VP4 gene sequence.

Lin et al, J Med Virol 78: 254, 2006


Evolution of enterovirus 71

Appearance of intratypic recombination of EV71

  • Recombination is a well-known phenomenon for enteroviruses.

  • The actual extent of recombination within circulating EV71 is not known.


Evolution of enterovirus 71

Genetic analysis of EV71


Evolution of enterovirus 71

1986

1998

1999

2000

2001

2002

2003

2004

2005

CA16

A

BrCr

7423-MS-87

72

237-TW-86

Phylogenetic analysis of EV71 isolates in Taiwan from 1986 and 1998-2005 based on VP1 region

242-TW-86

99

99

N2838-TW-03

9-97-SHA89

99

5145-TW-98

236-TW-86

97

99

N3793-TW-01

99

N0991-TW-01

S0320-TW-02

81

B

N2443-TW-02

N3496-TW-02

5666-SIN-002209

99

91

5865-SIN-000009

99

N0781-TW-01

N1643-TW-00

H0148-TW-00

80

S0875-TW-99

S0942-TW-99

S0943-TW-99

99

M0167-TW-00

N0716-TW-00

M0201-TW-00

99

S18191-SAR-02

02363-Thai-02

93

1814-JAP-01

1585-JAP-01

99

99

1695-JAP-01

2086-TW-98

99

5746-TW-98

H0001-TW-98

H0358-TW-98

NCKU9822

6092-TW-98

99

H0139-TW-98

4643-TW-98

1245a-TW-98

4-SHZH-01

C

2779-JAP-02

N3340-TW-02

SHZH03

99

40-SHZH-02

03-SHZH-01

08-SHZH-01

99

62-SHZH-02

N2735-TW-04

S0584-TW-04

N2121-TW-05

99

N2472-TW-04

N1862-TW-05

N2490-TW-04

N1614-TW-04

N0003-TW-05

N3031-TW-04

N2411-TW-04

0.1

0.1


Evolution of enterovirus 71

CA16

A

BrCr

NCKU9822

6092-TW-98

2086-TW-98

5746-TW-98

H0001-TW-98

H0139-TW-98

H0358-TW-98

4643-TW-98

1245a-TW-98

SHZH03

99

C

N3340-TW-02

N0003-TW-05

N1862-TW-05

N2735-TW-04

S0584-TW-04

N2490-TW-04

N3031-TW-04

N2411-TW-04

N2472-TW-04

N1614-TW-04

N2121-TW-05

237-TW-86

99

242-TW-86

236-TW-86

7423-MS-87

N2838-TW-03

5145-TW-98

93

9-97-SHA89

99

N2443-TW-02

99

N0991-TW-01

N3793-TW-01

B

S0320-TW-02

N3496-TW02

89

5666-SIN-002209

5865-SIN-000009

98

N1643-TW-00

N0781-TW-01

91

M0201-TW-00

N0716-TW-00

M0167-TW-00

S0875-TW-99

H0148-TW-00

S0942-TW-99

S0943-TW-99

0.1

Phylogenetic analysis of EV71 isolates in Taiwan from 1986 and 1998-2005 based on 3D region

1986

1998

1999

2000

2001

2002

2003

2004

2005


Evolution of enterovirus 71

CA16

A

BrCr

2086-TW-98

99

H0358-TW-98

H0139-TW-98

4643-TW-98

C

NCKU9822

1245a-TW-98

6092-TW-98

5746-TW-98

H0001-TW-98

N3340-TW-02

83

81

SHZH03

99

N0003-TW-05

N2121-TW-05

99

N1862-TW-05

B

S0584-TW-04

1986

N2735-TW-04

N2472-TW-04

1998

N2490-TW-04

1999

N2411-TW-04

99

2000

N3031-TW-04

N1614-TW-04

2001

7423-MS-87

242-TW-86

2002

99

236-TW-86

2003

237-TW-86

2004

9-97-SHA89

91

5145-TW-98

2005

N2838-TW-03

N0991-TW-01

N3793-TW-01

92

99

S0320-TW-02

B

N3496-TW-02

N2443-TW-02

N0781-TW-01

N1643-TW-00

5666-SIN-002209

5865-SIN-000009

M0201-TW-00

N0716-TW-00

S0942-TW-99

S0943-TW-99

S0875-TW-99

H0148-TW-00

M0167-TW-00

0.1

Phylogenetic analysis of EV71 isolates in Taiwan from 1986 and 1998-2005 based on 2B region


Evolution of enterovirus 71

Year

T

otal EV71

EV71 analyzed

Genotype of EV71 tested based on five regions

isolated

in this study

B

C

Genotype of

Recombinant

no. (%)

no. (%)

no (%)

5'

-

UTR

VP1

VP4

-

VP2

2B

3D

1986

unknown

7

7

(100.0)

0

0

B

B

B

B

B

1998

132

10

2 (20.0)

0

B

B

B

B

B

8 (80.0)

C

C

C

C

C

1999

13

4

4

(100.0)

0

0

B

B

B

B

B

2000

171

15

15

(100.0)

0

0

B

B

B

B

B

2001

116

5

5

(10

0.0)

0

0

B

B

B

B

B

2002

25

8

7 (100.0)

0

B

B

B

B

B

1 (12.5)

C

C

C

B

C

2003

10

4

4

(100.0)

0

0

B

B

B

B

B

2004

32

12

0

0

12

(100.0)

C

C

C

B

C

2005

34

8

0

1 (12.5)

C

C

C

C

C

7 (87.5)

C

C

C

B

C

Recombination of EV71 from 1986 and 1998-2005


Evolution of enterovirus 71

3B

5′UTR

3′UTR

VP2

VP3

VP4

2A

2C

3A

3C

3D

2B

VP4

Simplot compared N3340/TW/02 with EV71 reference strains


Evolution of enterovirus 71

3B

5′UTR

3′UTR

VP2

VP3

VP4

2A

2C

3A

3C

3D

2B

VP4

Bootscan compared N3340/TW/02 with EV71 reference strains


Evolution of enterovirus 71

Summary of EV71 recombination study

  • Intratypic recombination of EV71 among nature circulating isolates was found to be initiated in 2002 and became predominant in 2004 and 2005.

  • Recombinant points were located at the 3'-termini of 2A and 3C regions, suggesting that nonstructural genes recombined to produce new EV71 variants.


Evolution of enterovirus 71

Implications of EV71 study

  • The results indicated that standard typing may produce misleading results or even sequencing of multiple partial regions of EV71 will not always be sufficient.

  • It remains to be determined what role of the emergence of this new EV71 recombinant plays (e.g. clinical significance, vaccine selection).


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