Foot and mouth disease
OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / Foot and mouth diseaseAddress 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: / www.chvst.com
Name (including Title and Position) of Head of Laboratory (Responsible Official): / Dr Hong Yin, Head of Institute
Name(including Title and Position) of OIE Reference Expert: / Dr Xiangtao Liu, Director of Laboratory
Name (including Title and Position) of writer of this report
(if different from above): / Dr Jianhong Guo, Deputy Director
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
Test / For / Specificity / TotalLPB-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 / 25Suckling 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
2. 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
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.
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.
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 / Strain28/02/2011 / 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
07/04/2011 / FMD / O / Cattle / 252 / 87 / 12 / 240 / Jing Xiang village, Tianzhu, Qian dongnan , GUIZHOU / PanAsia
Sheep / goats / 7 / 0 / 0 / 7
Swine / 421 / 4 / 2 / 419
21/07/2011 / FMD / O / Cattle / 73 / 46 / 0 / 73 / Longfeng village, Pudi, Bijie, GUIZHOU / PanAsia
Swine / 124 / 78 / 0 / 124
Sheep / goats / 197 / 9 / 0 / 197
05/09/2011 / FMD / O / Cattle / 1744 / 6 / 6 / 1738 / Liebugou, Lengda village, Jiacha, Shannan, TIBET / PanAsia
10/10/2011 / FMD / O / Cattle / 108 / 7 / 0 / 108 / Duixu village, Zhongda town, Lang, Linzhi, TIBET / PanAsia
Sheep / goats / 2 / 0 / 0 / 2
Swine / 22 / 0 / 0 / 22
17/10/2011 / FMD / O / Cattle / 389 / 26 / 0 / 389 / Shuangjing village, Haiyuan, Zhongwei, NINGXIA / PanAsia
Sheep / goats / 293 / 0 / 0 / 293
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 (PCP-FMD) 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 / Organizer19-22/07 / 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
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, DrQiang 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.