OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Lanzhou Veterinary Research Institute, CAAS
Xujiaping No.1, Yanchangpu
Lanzhou,
Gansu province
Post code: 730046
THE PEOPLE’S REPUBLIC OF CHINA
Tel.:
+86-931-8342585
Fax:
+86-931-8340977, +86-931-8342052
e-mail address:
website:
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Name(including Title and
Position) of OIE Reference
Expert:
Name (including Title and
Position) of writer of this report
(if different from above):
Annual reports of OIE Reference Centres, 2011
Foot and mouth disease
hnxiangtao@hotmail.com; gregjh@126.com
www.chvst.com
Dr Hong Yin, Head of Institute
Dr Xiangtao Liu, Director of Laboratory
Dr Jianhong Guo, Deputy Director
1
Foot and mouth disease
Part I: Summary of general activities related to the disease
1.
Test(s) in use/or available for the specified disease/topic at your laboratory
The China National Foot-and-Mouth Disease Reference Laboratory (CNFMDRL) at LVRI is a central facility in
China for FMD diagnosis, while most monitoring samples were tested by provincial animal CDC. Tests used for
detection of viral antigen, antibodies and viral genome and the number listed below just were done in CNFMDRL
in 2011 (Table 1).
Table 1
2.
Test
For
Specificity
Total
LPB-ELISA
Antibody
Type O, Asia1 and A
6930
SPC-ELISA
Antibody
Type O, Asia1 and A
1050
VN
Antibody
Isolations
25
NSP-3ABC-ELISA
Antibody
DIVA
6930
IS-ELISA
Antigen
Type O, Asia1 ,A and SVDV
53
RIHA
Antigen
Type O, Asia1,A
53
BHK-21 cell culture
Virus isolation
Epidemic isolations
25
Suckling mice
Virus isolation
Epidemic isolations
53
RT-PCR
(including multiple RT-PCR , real-time
RT-PCR, and typing RT-PCR)
Molecular test
Group or Type
5020
VP1 sequencing
Molecular analysis
Epidemic Strain
49
Production and distribution of diagnostic reagents
Five types of reagent for FMD diagnosis were produced in-house officially, all of which were obtained new drug
certificate approved by Ministry of Agriculture, P.R China. The factory has been licenced GMP and ISO9001
international quality management system certification in 2011. The reagents/ kits produced were enough for
domestic demands; however, no reagent or kit was supplied to other countries’ laboratory. The type of reagent and
the amount supplied nationally were shown in Table 2.
Table 2
2
Type of reagent
Amount supplied nationally (including for own use)
LPB-ELISA Kit
~8000 kits (3000 kits for type O, 3000 kits for Asia1 and 2000 kits for type A)
NSP-3ABC-ELISA kit
~900 kits
IHA Antigen(for type O)
45,000 ampoules of 5ml for each, which can detect 11,250,000 serum specimens.
multiple RT-PCR Kit
~500 kits used for 10,000 suspected samples.
IS-ELISA kit
~20 kits used only in the CNFMDRL.
Annual reports of OIE Reference Centres, 2011
Foot and mouth disease
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
International harmonisation and standardisation of methods for diagnostic testing or the
production and testing of vaccines
a)
Establishment and maintenance of a network with other OIE Reference Laboratories
designated for the same pathogen or disease and organisation of regular inter-laboratory
proficiency testing to ensure comparability of results
Not done.
b)
Organisation of inter-laboratory proficiency testing with laboratories other than OIE
Reference Laboratories for the same pathogens and diseases to ensure equivalence of
results
Not done.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
The standard reference materials in our Laboratory are not officially recognised by OIE or other international
bodies. These materials are all made and identified in our Laboratory using molecular and serological methods.
The reference sera are available for serotypes O, A and Asia 1. Pigs or cattles serum against O/Mya/98,
O/Tibet/99, Asia 1/JS/CHA/05, A/F72 (A22 lineage) and A/WH/09 (SEA topotype) are available for serology
tests and other studies. We can also provide the inactivated antigen of above isolates.
5.
Research and development of new procedures for diagnosis and control
5.1 Techniques Developed for Foot-and-Mouth Disease Diagnosis in CNFMDRL
5.1.1 Development and validation of a lateral flow immunoassay using colloidal gold for the identification of
serotype-specific foot-and-mouth disease virus O, A and Asia 1[see J. Virol. Methods, 2011, 171: 74-80.]
5.1.2 2C`3AB-ELISA for detection of antibodies against NSP
5.1.3 Development of a dot immunoblot method for differentiation of animals infected with foot-and-mouth
disease virus from vaccinated animals using non-structural proteins expressed prokaryotically [see J. Virol.
Methods, 171 (2011) 234–240]
5.1.4 Detection of foot-and-mouth disease virus RNA by reverse transcription loop-mediated isothermal
amplification.
5.2 New Vaccine Developments of Foot-and-Mouth Disease in CNFMDRL
5.2.1 Engineering Foot-and-Mouth Disease Viruses with Improved Growth and Protective Potency for
Vaccine.
A novel plasmid-based reverse genetics system was used to construct a chimeric strain by replacing the P1 gene in
the vaccine strain O/CHA/99 with that from the epidemic stain A/WH/CHA/09. The potency of the rA/P1-FMDV
antigens was 10.81, 13.59 and 13.59 PD50 per dose (3 μg) in three tests; the potency of the A/WH/CHA/09
antigens was 5.27 PD50 per dose (3 μg).The recombinant vaccine has been authorized to manufacture by the
Vaccine Evaluation Committee of Ministry of Agriculture in China.
5.2.2 Selection or Engineering O type Foot-and-Mouth Disease Viruses with Broaden Coverage of Antigen
for the Development of Improved Vaccine
After the outbreak affected with O/Mya-98 strain, a new vaccine strain (Re-O/Mya-98) had been constructed
successfully by LVRI last year. The vaccine strain has broad coverage of antigen and high cross-protection against
PanAsia, Mya-98 and Cathay strains. The vaccine has been authorized to manufacture by Ministry of Agriculture
in China.
Annual reports of OIE Reference Centres, 2011
3
Foot and mouth disease
5.2.3 FMDV RGS (reverse genetics system)-based DNA vaccine (continue to do)
5.2.4 Promising Multiple-Epitope Recombinant Vaccine against Foot-and-Mouth Disease Virus Type O in
Swine. Clinical And Vaccine Immunology[see Clinical And Vaccine Immunology, 2011, 18(1): 143-149]
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
There are 7 FMD outbreaks were confirmed and reported to OIE immediately in 2011. 596 animals including
swine, cattle and sheep/goat were infected, and 7733 animals were killed. The main epidemic virus is type O
FMDV, belonged SEA topotype Mya-98 lineage (n=2) and ME-SA topotype PanAsia strain (n=5). One epidemic
PanAsia strain collected from Guizhou Province VP1 sequence was submitted to GeneBank (Accession Number is
JF837375). These viruses were very closely related to recent viruses from SEA nations. No type A and Asia1
outbreaks and positive samples were found this year (Table 3).
From 15th April to 15th June 2011, an active, routine and large-scale epidemiological survey was organized and
implemented with China Animal Disease Center under direction of Veterinary Bureau of Ministry of Agriculture.
The monitoring activity covered seven provinces (Yunnan, Guangxi, Guizhou, Guangdong, Hunan, Sichuan and
Chongqing) in southwest of China and received some valuable results for FMD prevention and control in above
zones. In addition, four emergency epidemiological surveys were carried out in Xinjiang Autonomous Region,
Guizhou Province and Tibet after FMD outbreaks this year. In 2012, a systematic epidemiological survey will be
held across the border area in Xinjiang Autonomous Region.
Quarterly monitoring for Hainan Island FMD-free zone with immunization was kept implementing.
Table 3
Report
Date
Disease
Type
Species
Susceptible
Cases
Deaths
Destory
Location
Strain
28/02/2011
FMD
O
Swine
3941
275
0
3941
Qia'erbagexiang, Kuerle, Bazhou,
XINJIANG
mya-98
28/03/2011
FMD
O
Swine
205
58
25
180
Xinjiang Production and
Construction Corps, XINJIANG
mya-98
Cattle
252
87
12
240
Sheep /
goats
7
0
0
7
Jing Xiang village, Tianzhu, Qian
dongnan , GUIZHOU
PanAsia
Swine
421
4
2
419
Cattle
73
46
0
73
Swine
124
78
0
124
Longfeng village, Pudi, Bijie,
GUIZHOU
PanAsia
Sheep /
goats
197
9
0
197
Cattle
1744
6
6
1738
Liebugou, Lengda village, Jiacha,
Shannan, TIBET
PanAsia
Cattle
108
7
0
108
Sheep /
goats
2
0
0
2
Duixu village, Zhongda town, Lang,
Linzhi, TIBET
PanAsia
Swine
22
0
0
22
Cattle
389
26
0
389
Shuangjing village, Haiyuan,
Zhongwei, NINGXIA
PanAsia
07/04/2011
21/07/2011
05/09/2011
10/10/2011
17/10/2011
FMD
FMD
FMD
FMD
FMD
O
O
O
O
O
Sheep /
goats
4
293
0
0
293
Annual reports of OIE Reference Centres, 2011
Foot and mouth disease
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
7.1On 11th, November, 2011, China National Accreditation Service for Conformity Assessment (CNAS ）
transvalued the ABSL-3 Laboratory of CNFMDRL according to the new accreditation criteria. CNAS considered
that the Laboratory meets the relevant requirements of CNAS-CL05:2009 (Accreditation Criteria for Biosafety
Laboratories), give the ABSL-3 accreditation certificate (the certificate No. is CNAS BL0012).
7.2 The construction of the CNFMDRL manufacture facility for diagnostic reagents has been completed and
passed the certificate of the quality certification system of ISO 9001 and GMP within 2011.
8.
Provision of consultant expertise to OIE or to OIE Member Countries

Participation in OIE working group/meeting(see item 13)

CNFMDRL submitted amendments and suggestions to the Progressive Control Pathway for FMD control (PCPFMD) and the report of the scientific commission for animal diseases of OIE. We welcome the PCP-FMD been
adopted by FAO and OIE as a working tool in the design of FMD control programs, we suggest that the
specialized work committee draw up the valuation sheets of different stage according to PCP principle, provides
its to the countries (zones) for valuating themselves and control FMD .
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
Date
Name of workshops,
seminars, technical training
courses
Host place
No. of Attendees/
Country come from
Organizer
19-22/07
FAO assistance DPRK Program,
training on laboratory diagnostic
techniques of FMD
Pyongyang,
DPRK
10/DPRK
FAO,CNFMDRL
15-25/10
Workshop on FMD Control
Technologies for CAREC
Countries
Beijing, China
15/ Mongolia,
Kazakhstan, Tajikistan,
Kyrghizstan,Uzbekistan
MoA of China,
CNFMDRL
10. Provision of diagnostic testing facilities to other OIE Member Countries
In 2011, CNFMDRL detected two tissues samples from DPRK and Vietnam. The virus from DPRK is belonged to
serotype O, SEA topotype Mya-98 strain, and the sample collected from Vietnam is O/ME-SA/PanAsia strain.
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
No.
12. Participation in international scientific collaborative studies
12.1
IAEA project, Part of the IAEA’s Coordinated Research Project (CRP)
12.2
EPIZONE project WP6.3 (Experimental Epidemiology) IC 4.7 and molecular epidemiology of foot
and mouth disease virus in Asia.
12.3
FMD-DISCONVAC(226556): Development, enhancement and complementation of animal-sparing,
foot-and-mouth disease vaccine-based control strategies for free and endemic regions
Annual reports of OIE Reference Centres, 2011
5
Foot and mouth disease
13. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)

Presentations at international conferences and meetings
 Dr Jijun He participated in the Joint SEACFMD Laboratory Network (LabNet) and Epidemiology Network
(EpiNet) Meeting hold in Pak Chong, Thailand from March 2 to 3, 2011，and gave a presentation entitled
‘National FMD Reference Laboratory Recent 5-Year Report’.
 Professor Hong Yin, Director General of Lanzhou Veterinary Research Institue, led a delegation to participate
in the 5th International Meeting of Epizone hold in Netherland from April 11 to 14, 2011. In this meeting,
Dr Qiang Zhang and Dr Haixue Zheng from CNFMDRL gave speechs to introduce the research development
in our laboratory.
 Dr Huiyun Chang participated in the Expert Forum on FMD hold by International Atomic Energy Agency
(IAEA）in Vienna, Austria from April 17 to 22, 2011. In this meeting, Dr Chang introduced the situation of
FMD control in China and discussed some techinical issues on FMD contorl.
 Dr Jianhong Guo representing CNFMDRL as a member of Chinese delegate visited Democratic People's
Republic of Korea (DPRK) to support the FMD control effort in this country, and discussed the common
concerned O/Mya/98 virus prevalence in China and DPRK in May, 2011.
 Dr Qiang Zhang visited DPRK invited by FAO as a FMD expert to conduct the training workshop on FMD
diagnosis in June, 2011.
 Dr Haixue Zheng was invited to give a presentation entitled ‘reverse genetic operation technique and its usage
in FMDV research’ in China-EU Youth Symposium hold in Beijin in July, 2011.
 Dr Shiqi Sun and Haixue Zheng participated in the OIE/WHO SEACFMD Laboratory Director Meeting hold
in Kuala Lumpur, Malaysia in Oct. 18, and the Meeting on Development of Laboratory Capability for
Diagnosis of New Emerging Infectious Disease from Oct. 19 to 21, 2011.
 Dr Zengjun Lu, as a member of Chinese delegate, participated in the 27th Conference of the OIE Regional
Commission for Asia, the Far East and Oceania hold in Tehran, Iran in December, 2011.
 Dr Shiqi Sun and Haixue Zheng participated in the meeting on FMD control hold in Tokyo in Nov.1, 2011,
and gave a presentation entitled FMD Control and Eradication Strategies in China’.

Scientific publications in peer-reviewed journals
At least 20 articles were collected in SCI paper written in English and more than 70 papers were published in
Chinese journals. The SCI papers were list below.
 Du J, Gao S, Luo J, Zhang G, Cong G, Shao J, Lin T, Cai X, Chang H. Effective inhibition of foot-and-mouth
disease virus (FMDV) replication in vitro by vector-delivered microRNAs targeting the 3D gene. Virol J,
2011, 8: 292.
 Shao JJ, Wong CK, Lin T, Lee SK, Cong GZ, Sin FWY, Du JZ, Gao SD, Liu XT, Cai XP, Xie Y, Chang HY,
Liu JX. Promising Multiple-Epitope Recombinant Vaccine against Foot-and-Mouth Disease Virus Type O in
Swine. Clinical And Vaccine Immunology, 2011, 18(1): 143-149.
 Luo JH, Du JZ, Gao SD, Zhang GF, Sun JJ, Cong GZ, Shao JJ, Lin T, Chang HY. Lentviral-mediated RNAi to
inhibit target gene expression of the porcine integrin av subunit, the FMDV receptor, and against FMDV
infection in PK-15 cells. Virol J, 2011, 8: 428.
 Lin T, Shao JJ, Du JZ, Con GZ, Gao SD, Chang HY. Development of a serotype colloidal gold strip using
monoclonal antibody for rapid detection type Asia1 foot-and-mouth disease. Virol J, 2011, 8: 418.
 Jiang T, Liang Z, Ren WW, Chen J, Zhi XY, Qi GY, Yang YM, Liu ZX, Liu XT, Cai XP. Development and
validation of a lateral flow immunoassay using colloidal gold for the identification of serotype-specific footand-mouth disease virus O, A and Asia 1. J Virol Meth, 2011, 171: 74-80.
6
Annual reports of OIE Reference Centres, 2011
Foot and mouth disease
 Li PH, Lu ZJ, Bao HF, Li D, King DP , Sun P, Bai XW, Cao WJ, Gubbins S, Che YL, Xie BX, Guo JH, Yin
H, Liu ZX. In-vitro and in-vivo phenotype of type Asia 1 foot-and-mouth disease viruses utilizing two nonRGD receptor recognition sites. BMC Microbiology, 2011, 11: 154.
 Wu L, Jiang T, Lu ZJ, Yang YM, Sun P, Liang Z, Li D, Fu YF, Cao YM, Liu XT, Liu ZX. Virol J, 2011, 8:
186. Development and validation of a prokaryotically expressed foot-and-mouth disease virus nonstructural
protein 2C’3AB-based immunochromatographic strip to differentiate between infected and vaccinated animals.
Virol J, 2011, 8: 186.
 Cao YM, Lu ZJ, Sun P, Fu YF, Tian FP, Hao XF, Bao HF, Liu XT, Liu ZX. A pseudotype baculovirus
expressing the capsid protein of foot-and-mouth disease virus and a T-cell immunogen shows enhanced
immunogenicity in mice. Virol J, 2011, 8: 77.
 Fu YF, Cao YM, Sun P, Bao HF, Bai XW, Li PH, Li D, Lu ZJ, Liu ZX. Development of a dot immunoblot
method for differentiation of animals infected with foot-and-mouth disease virus from vaccinated animals
using non-structural proteins expressed prokaryotically. J Virol Meth, 2011, 171: 234-240.
 Bai XW, Li PH, Bao HF, Liu ZX, Li D, Lu ZJ, Cao YM, Shang YJ, Shao JJ, Chang HY, Luo JX, Liu XT.
Evolution and molecular epidemiology of foot-and-mouth disease virus in China. Chinese Science Bulletin,
2011, 56(21): 2191-2201.
 Li ZY, Liu JX. The current state of vaccines used in the field for foot and mouth disease virus in China. Expert
Rev. Vaccines, 2011, 10(1): 13-15.
 Li ZY, Yin XP, Yi YZ, Li XR, Li BY, Lan X, Zhang ZF, Liu JX. FMD subunit vaccine produced using a
silkworm-baculovirus expression system: protective efficacy against two type Asia1 isolates in cattle. Vet
Microbiol, 2011, 149(1-2): 99-103.
 Zhang L, Zhang J, Chen HT, Zhou JH,Ma LN, Ding YZ, Liu YS. Research in advance for FMD Novel
Vaccines. Virol J, 2011, 8: 268.
 Ma LN, Zhang J, Chen HT, Zhou JH, Ding YZ, Liu YS. An overview on ELISA techniques for FMD. Virol J,
2011, 8: 419.
 Liu XS, Wang YL, Zhang YG, Fang YZ, Pan L, Lu JL, Zhou P, Zhang ZW, Jiang ST. Identification of H-2d
Restricted T Cell Epitope of Foot-and-mouth Disease Virus Structural Protein VP1. Virol J, 2011, 8: 426.
 Liu Y, Zhang K, Zheng H, Shang Y, Guo J, Tian H, Lu G, Jin Y, He J, Cai X, Liu X. Proteomics Analysis of
Porcine Serum Proteins by LC-MS/MS after Foot-and- Mouth Disease Virus (FMDV) Infection. J Vet Med
Sci. 2011 Jul 29.
 Zhou JH, Zhang J, Chen HT, Ma LN, Ding YZ, Pejsak Z, Liu YS. The codon usage model of the context
flanking each cleavage site in the polyprotein of foot-and-mouth disease virus. Infect Genet Evol. 2011 Jul 23.
 Zheng H, He J, Guo J, Jin Y, Yang F, Lv L, Liu X. Genetic characterization of a new pandemic Southeast Asia
topotype strain of serotype O foot-and-mouth disease virus isolated in China during 2010. Virus Genes, 2011
Sep 20.
 Bao HF, Li D, Sun P, Zhou Q, Hu J, Bai XW, Fu YF, Lu ZJ, Liu ZX. The infectivity and pathogenicity of a
foot-and-mouth disease virus persistent infection strain from oesophageal-pharyngeal fluid of a Chinese cattle
in 2010. Virol J, 2011, 8(1):536.
 Wang G, Pan L, Zhang Y, Wang Y, Zhang Z, Lü J, Zhou P, Fang Y, Jiang S. Intranasal delivery of cationic
PLGA nano/microparticles-loaded FMDV DNA vaccine encoding IL-6 elicited protective immunity against
FMDV challenge. PLoS One. 2011;6(11):e27605.

Other communications
On 19th SEP, 2011, the thirteenth China FMD Scientific Research Conference was hold in Huangshan, Anhui
Province. Around 300 people from provincial animal CDC, research institutes, and universities attended the
meeting. More than 20 staffs from our lab were involved in the discussion and made reports related to FMD
Epidemology, Vaccine and Vaccination program, Prevention and Control. The meeting organized by the FMD
Branch under Chinese Association of Animal Service and Veterinary Medicine, which is an important platform for
FMD researchers in China.
Annual reports of OIE Reference Centres, 2011
7