2009,25(18):457-460

Chinese Agricultural Science Bulletin

266109

FG-9307

2





S941.1 A
2009-1477

Study of Lymphocystis Disease Virus Infected to Flounder Gill Cell Line

Cheng Shunfeng
(Qingdao Agriculture University, Qingdao Shandong 266109)
Abstract: In this article, the nodule of the diseased flounder Paralichthys olivaceus with lymphocystis disease
was collected, and lymphocystis disease virus (LCDV) was separated from lymphocystis cells of nodule by cell
disruption, centrifugation. Flounder gill cell line (FG-9307) was used to confirm susceptive to LCDV. After
passed through filter, the virus suspension was inoculated with FG-9307 cells cultured in 96-well plates. 1~2
days later, FG cells exhibit cytopathic effect (CPE). Indirect immunofluorescence assay test (IFAT) was
employed to detect viral infected by using monoclonal antibodies against LCDV. The result of IFAT was
positive which the FG-9307 cells inoculated with LCDV, and the positive signals occurred at the cytoplasm.
The result of IFAT was negative which FG cells not inoculated with virus suspension. These date proved that
FG-9307 cell line was susceptive to LCDV.
Key words: lymphocystis disease virus, FG-9307 cell line, cytopathic effect

0 [2] LCDV
Lymphocystis disease LCD [2-3] [4-5] [6] [7-8]
42 125
20 60
Lymphocystis disease virus, LCDV



[1]

Epinethelus moara BF-2LBF-1GF-1
Lateolabrax japonicus Lutjanus [3]
argentimaculatus FG-9307

Y2007D31, Y2007D74630701
1978 266109
Tel0532-86080734E-mailchengshunfeng@gmail.com
2009-07-02
2009-07-24
 458 http://www.casb.org.cn

4 PBS 30 l
LCDV
1.2.4
1 1 cm1 cm

1 2 6
1.1
FG 3 PE
1~2 4 0.5 ml
MEM/EBSS Hyclone 20 1 h 2 ml20
Trypsin 1:250 AMRESCO PBS 0.5 ml
4PBS
Coast FITC IgG 3
20 min
Sigma 5
37 45 min
PBS 3
5 min
1.2 6 FITC IgG
1.2.1 FG 1 L MEM/ 37 45 minPBS 3 5 min
EBSS NaHCO3 2.2 g 110 5
7


IU/L 110 g/L pH 7.2~7.4
5
2
4 2.1
2%

10% 3 (6~
7)105 cells/ml
(PE)0.01 MpH 7.4 1A
0.02% EDTA 2.2 FG
1.2.2 24 h
1 10 g CPE

2 10 TNE50 mM Tris100 mM
NaCl1 mM EDTApH 7.4 48h
10~20 s
3 CPE 1B
3 500g30 min
4 2.3
FG
4 4 1 800g 30 min
FG 2A
5 0.45 m LCDV FG
-20 2B
1.2.3 LCDV FG
1 FG 96 100 l FG 0 h
20 12 h
2PE 24 h 72 h
1~2
30 l 96h
320 1 h20
CPE
 459

A B

1
A
B

A B

2
A B
48h

3
[8]
Iwamoto 2002
39
HINAE
CPE Garca-Rosado
[3]

[8]
Zhang GCO
[9]
GCK
2 48 h
FG 72 h

48~72 h

1
2
30~60 min

pH [10] 60 min
3

4PCR
5

12 h
[10]
GarcaRosado
 460 http://www.casb.org.cn

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