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FOREIGN ANIMAL DISEASE DIAGNOSTIC LABORATORY OVERVIEW OF FISCAL YEAR 2016 KARYN A . HAVAS , DVM, PHD D IAGNOST IC SERV ICES SECT ION HEAD U.S . DEPARTMENT OF AGRICULTURE AN IMAL AND PLANT HEALTH INSPECT ION SERV ICE VETER INARY SERV ICES SC IENCE , TECHNOLOGY, AND ANALYS IS SERV ICES OCTOBER 18 , 2016
FY 2016 FAD InvesCgaCons & Surveillance
Priority
Level Accessions Final Diagnosis Domes5c
Disease Diagnosis
FAD ruled out / Dx Undetermined
SVA EHD PPox BT MCF VSV Total
A 1 -‐ -‐ -‐ -‐ -‐ -‐ 0 1
2 188 59 4 2 1 0 2 67 121
3 262 66 2 7 1 2 31 108 154
Total 451 125 6 9 2 2 33 177 274
CSF Surveillance
Accessions Samples Assays
1,952 8,463 8,475
SVA [PERCENTAGE]
VSV 48%
EHD 7%
MCF 3%
BT 0% PPOX
12%
FY15
3
FY 15 & 16 FAD Inves5ga5ons
SVA [PERCENTAGE
] VSV 19%
EHD 3%
MCF 1%
BT 1%
PPOX 5%
FY16
Assays Run in FY 2016 FAD accessions (n= 451) Assay Count
PCR 6,440 Ab ELISA 2,279 Complement Fixa5on 162 Virus isola5on 1,494 Electron microscopy 6 Histopathology 16 Agar gel immunodiffusion (AGID) 332 Immunoperoxidase (IP) 17 Direct Fluorescent An5body (DFA) 15 Indirect Fluorescent An5body (IFA) 266 Mycoplasma isola5on 0 Immunohistochemistry 46
TOTAL 11,073
4
DiagnosCc Reagents and Biologics Produced FY 16
Type of Material
# of Lots Produced
Quan5ty
Cultured Cells 112 14,570 flasks
Master Cell Stocks 3 1,511 vials
Macrophages 2 20
Culture Media 54 319,507 mL
Condi5on Media (L929) 1 3,300 mL
Solu5ons 5 73,200 mL
Monoclonal An5bodies 2 600 mL
An5sera 1 2,800 mL
Viruses 22 5,700 mL
Plates/Slides CSF IP Plates VESV slide ASFV slide
100 plates 172 slides 30 slides
Mycoplasma culture 2 2,000 mL
PrioCHECK panels 49 3,621 vials
BEI inac5va5on 16 3,900 mL
Other materials 28 910 vials 5
Number of Registered Labs and Par5cipants for FY 2016 PT Panels
CSF FMD ASF CSF/FMD FY16
Pre-‐test Rinderpest Virus Total
Number of registered labs 44 45 9 10 N/A 108
Number of par5cipants 201 207 53 3 N/A 464
Number of PT panels 1920 1920 800 NA 720 5360
Number of posi5ve amplifica5on controls
540 (combined) NA NA 321 861
Number of posi5ve extrac5on controls
540 (combined) NA NA 320 860
TOTAL 7081 6
PT PANELS AND REFERENCE MATERIALS DISTRIBUTED IN FY 16
CSF FMD ASF CSF/FMD FY15 PT Retest
CSF/FMD Surveillance Controls
FMD/ASF Surveillance Controls
Total
PT Panels Shipped 201 207 53 8 N/A N/A 469
Reference Panels Shipped 92 92 18 NA N/A N/A 110
Controls Shipped 454
(FMD/CSF)
106 NA 840 (CSF/FMD combined)
106 (ASF) 1506
Number of Shipments
50 (FMD/CSF Comb)
9 4 45 8 116
7
PT RESULTS ANALYZED AND SCORED IN FY16 NVSL-‐FADDL
CSF FMD ASF PT Results Analyzed and Scored in FY16 from FY15 PT (not completed in FY2015)
24 30 0
PT Results to be scored in FY17 from FY16 PT (est.) 201 207 53
FAD DEMONSTRATIONS IN FY16 (11 FAD demonstrated per course)
Course Number of Animals Prepared
Number of Inocula5ons
Foreign Animal Disease DiagnosScian Course (1) 132 122
Veterinary Laboratory DiagnosSc Course (1)
Area Foreign Animal Disease DiagnosScian Refresher Course (2)
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Training
9
Inac5vated Materials in FY15 -‐16
Safety Treatment Safety Tes5ng
2015 2016 2015 2016
69 samples
7 samples 13 imports (5 in vivo)
11 imports (2 in vivo)
VAC Tes5ng Conducted in the NAFMDVB FY16
Test VAC
PD50 3
Stability Test 1
AnSgenic Mass DeterminaSon (146S) 5
Developmental Projects FY 16
Serologic assay development ◦ FMD 3ABC dot blot confirmatory serologic test ◦ VSV Microgrant 2015
◦ Development of OIE Standard Liquid Phase Blocking ELISA (LPBE) and Solid-‐Phase CompeKKon ELISA (SPCE) for FMD Vaccine Matching and Serotype-‐Specific AnKbody TesKng ◦ FADDL-‐DHS IAA 2014-‐2016
◦ FMD 3D ELISA for DIVA detecKon of FMDV anKbody in swine ◦ FADDL DHS IAA 2014-‐2017
Advanced Molecular Detec7on Methods ◦ Sample target enrichment using magneKc beads, subtracKve hybridizaKon ◦ FADDL DHS IAA 2014-‐2016
◦ uForge consorKum for microbial forensics and genomics project
◦ MinIon applicaKon
Molecular diagnos7cs ◦ Pen-‐side MulKplex DetecKon of FMDV and Bovine Papular StomaKKs Virus by a Portable Microfluidics PCR System for Rapid Clinical Disease DifferenKaKon ◦ FADDL DHS IAA 2016-‐2017
◦ FMD Serum Banking ◦ FADDL-‐DHS IAA 2014-‐2016
◦ Lumpy Skin Disease rPCR ◦ Development and Full ValidaKon of Parapox rPCR (completed)
Valida7on work ◦ ASF real Kme-‐ PCR ◦ FADDL-‐DHS IAA 2014-‐2016
◦ Gap analysis ASF ELISA ◦ FADDL-‐DHS IAA 2014-‐2016
◦ Methods comparison RNA/DNA dual extracKon ◦ FADDL-‐DHS IAA 2014-‐2016
◦ Biorad vs ABI 7500 comparison ◦ VSV IgM/IgG double-‐sandwich ELISA
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InternaConal AcCviCes 2016 Capacity Building & DiagnosIc Support
Capacity building and diagnos5cs ◦ Peru: PRRS diagnosScs and characterizaSon ◦ Dominican Republic: CSF diagnosSc support ◦ Panama: Vesicular diseases diagnosSc capacity building ◦ Tanzania: CSF and PCV PCR transfer and diagnosis of ASF in slaughterhouses ◦ Uganda: Syndromic surveillance training with non-‐profit and development of a Quality Management Course
Interna5onal conferences ◦ FAO-‐OIE Technical MeeSng on InternaSonal Preparedness and Vaccine Strategic Reserves for Rinderpest (Rome, Italy; April 2016)
◦ FMD Reference Lab MeeSng (Brussels, Belguim; November 2015) ◦ COSALFA (Uruguay; April 2016) ◦ ASF/CSF Interlaboratory Comparison MeeSng (Hannover, Germany; June 2016) ◦ CaribVet (Havana, Cuba; August 2016)
NBAF TransiCon Developing the future workforce
◦ FADDL Program of Requirements (POR) ◦ NBAF Core Services
◦ ORISE Fellowships at FADDL (6) ◦ Pathways Summer Intern (1) ◦ Saul T. Wilson Scholar (1) ◦ Synergies with Bob Rowland’s laboratory (KSU) ◦ RotaSon for veterinary medicine students and pathology
residents in the FADD schools (2)
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PublicaCons, FY16 Bracht AJ, O’Hearn ES, Fabian AW, Barreke RW, Sayed A. Real-‐Sme reverse transcripSon PCR assay for detecSon of Senecavirus A in swine vesicular diagnosSc specimens. PLoS One. 2016; 11(1):e0146211.
Duque H, Naranjo J, Carrillo C, Burbano A, Vargas J, Pauszek L, Olesen I, Sanchez-‐Vazquez MJ, Cosivi O, Allene RM. ProtecSon induced by a commercial bivalent vaccine against Foot-‐and-‐Mouth Disease in 2010 field virus from Ecuador. Vaccine. 2016; 34(35):4140-‐4144.
Das A, Ward G, Lowe A, Xu L, Renshaw R, Dubovi E, Reising M, Jia W. Development and ValidaSon of a Highly SensiSve Real-‐Time Polymerase Chain ReacSon Assay for Rapid DetecSon of Parapoxviruses. J Vet Diag Inv. (in press)
Das A, Deng MY, Babiuk S, McIntosh MT. ModificaSon of two capripoxvirus real-‐Sme PCR assays to include beta-‐acSn as an internal posiSve control to improve diagnosSc sensiSvity. (Submi^ed to JVM)
Lemire KA, Rodriguez YY, McIntosh MT. Alkaline hydrolysis to remove potenSally infecSous viral RNA contaminants from DNA. Virol J. 2016; 13:88.
Rhyan J, McCollum M, Gidlewski T, Shalev M, Ward G, Donahue B, Artz J, Stenfeldt C, Mohamed F, Nol P, Deng M, Metwally S, McKenna T, Salman M. Foot-‐and-‐mouth disease in a small sample of experimentally infected pronghorn (AnSlocapra americana). J Wildl Dis. 2016: [Epub ahead of print]
Sanford B, Holinka LG, O'Donnell V, Krug PW, Carlson J, Alfano M, Carrillo C, Wu P, Lowe A, Risam GR, Gladue DP, Borca MV (2016). DeleSon of the thymidine kinase gene induces complete anenuaSon of the Georgia isolate of African swine fever virus. Virus Res. 2; 213:165
Schuna C, Barrera J, Pisano M, Zsak L, Grubman MJ, Mayr GA, Moraes MP, Kamicker BJ, Brake DA, Enyreddy D, Brough DE, Butman BT, Neilan JG. MulSple efficacy study of an adenovirus-‐vectored foot-‐and-‐mouth disease virus serotype A24 subunit vaccine in canle using homologous challenge. Vaccine. 2016; 34(27): 3214-‐3220.
Shi X, Liu X, Wang Q, Das A, Ma G, Xu L, Sun Q, Peddireddi L, Jia W, Liu Y, Anderson G, Bai J, Shi J. A mulSplex real-‐Sme PCR panel assay for simultaneous detecSon and differenSaSon of 12 common swine viruses. J Virol Methods. 2016 Oct; 236:258-‐65.
Thomas LF, Bishop RP, Onzere C, McIntosh MT, Lemire KA, de Glanville WA, Cook EA, Fevre EM. Evidence for the presence of African swine fever in an endemic region of Western Kenya in the absence of any reported outbreak. BMC Vet Res. 2016; 12(1): 192.
Velazquez-‐Salinas L, Risam GR, Holinka LG, O’Donnell V, Carlson J, Alfano M, Rodriguez KK, Carrillo C, Gladue DP, Borca MV. Recoding structural glycoprotein E2 in classical swine fever virus (CSFV) produces complete virus anenuaSon in swine and protects infected animals against disease. Virology. 2016; 4949:178-‐189.
Xiju Shi, Liu X, Wang Q, Das, A, Ma G, Xu, L, Sun Q, Peddireddi L, Jia, W, Liu Y, Anderson G, Jianfa Bai, J, Jishu Shi, J (2016). A mulSplex real-‐Sme PCR panel assay for simultaneous detecSon and differenSaSon of 12 common swine viruses. J. Virol. Methods. 236; 258:265.
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QuesSons?
