FMDv NSP Ab Rapid test kit
Transcript of FMDv NSP Ab Rapid test kit
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Passion, Integrity, Professional, and Ethics
Excelsior Bio-System
P r o fe s s i o n a l a n d Re l i a b l e
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Profe s s i ona l and Re l i ab l e
Costs of eradicating 1997 outbreak estimated to be US$ 378.9 million,
Potential export losses of approximately US$ 1.2 billion (Data analysis Yang et al. (1999))
Foot & Mouth Disease (FMD) – A big economic Threat
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Profe s s i ona l and Re l i ab l e
Sentinel® FMDV Non Structural Protein Antibody Rapid Test
Sentinel® FMDV Non Structural Protein Antibody ELISA kit
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What is NSP?
4
Foot and mouth disease virus
Viral structural proteins, SP
Viral non structural proteins (NSPs)
Pro fe s s i ona l and Re l i ab l e
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Vaccine and viral protein
Viral structural proteins
Viral non structural proteins (NSPs)
Early vaccine
New vaccine
Not present inFMDV structure,Not in New vaccine!
Could be present
Only appeared in viral replication process
Profe s s i ona l and Re l i ab l e
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Removing NSP in vaccines
Distinguish between animals that have been vaccinated and animals that are infected!
FMD Vaccine (NSP removed)
inoculation
blood
Antibody : OnlyAnti-structural protein
infection
FMDV
Antibody : Anti-structural protein & Anti-nonstructural protein
blood
Profe s s i ona l and Re l i ab l e
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Differentiation Infected and Vaccinated Animal
Results serology
FMDV NSP test
Vaccinated Infected +
Vaccinated Not infected -Not
vaccinatedInfected +
Not vaccinated
Not infected -
P r o fe s s i o n a l a n d Re l i a b l e
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To prove absence of infection or virus circulation
To certify individual animal free of FMD prior to move or trade
To confirm suspected case
To distinguish from infected and vaccinated animal (DIVA)
NSP Ab testing for FMD are performed in support for 4 main purposes
Profe s s i ona l and Re l i ab l e
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Vaccinated or infected animals
Extremely Easy
High specificity and sensitivity
Monitor FMD Virus activity
Time saving
Profe s s i ona l and Re l i ab l e
Sentinel® FMDV NSP Ab Raipid test
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ELISA
Sentinal® FMDV NSP Ab Rapid Test
16 hours 60 mins 20 mins
Read by ELISA ReaderStop reactionWash
Add Secondary Antibody Develop with Substrate
Wash
Add Sample and Control
30 mins
Add Running buffer
Interpret by eyesAdd Sample
Over 17 hours!!
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Profe s s i ona l and Re l i ab l e
Protocol
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P r o fe s s i o n a l a n d Re l i a b l e
Validation test and Field test
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Profe s s i ona l and Re l i ab l e
• Swine sera provided by Animal Health Research Institute (AHRI).
• Total 62 positive swine samples from pigs experimentally infected
with FMDV O/Taiwan/1997 strain at 14 days post-infection(dpi)
were tested, and results are as follows:
Sensitivity :98.39 % (61/62)
Lab tests-Sensitivity
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P r o fe s s i o n a l a n d Re l i a b l e
Lab tests-Specificity
• Animal Health Research Institute (AHRI) provided swine sera and analyzed.
• Total 412 FMD negative swine sera from vaccinated pigs, pre-outbreak pigs and SPF pigs were tested.
Specificity :92.96 % (383/412)
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P r o fe s s i o n a l a n d Re l i a b l e
•A serum panel collected sequentially from 32 secondary SPF pigs
experimentally challenged by the FMDV O/Taiwan/1997 strain was
tested. The sera in the panel were sampled at different days post
infection(dpi).
•Results were compared with PrioCHECK, UBI and Chekit ELISA kit.testing limited
Tes
t p
osit
ive
%Lab tests-early testing
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The validation test was executed by the Animal Technology Institute Taiwan (ATIT).
169 swine sera were blind tested.Then compared results with PrioCHECK® FMDV NS (Ceditest)
and UBI® FMDV NS ELISA(swine), respectively.The data had showed that the Kappa value of Sentinel were
more than 0.8, it indicate the data of Sentinel® were almost the same result with PrioCHECK® and UBI®.
P r o fe s s i o n a l a n d Re l i a b l e
Observed agreement
Kappa value Sensitivity Specificity
PrioCHECK 0.95 0.87 90.2 % 96.9 %
UBI 0.95 0.86 86.4 % 97.6 %
Field tests-comparison with other kit
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Kappa( K coefficient of agreement; K)
P = (observed agreement):
P (chance agreement):=
─────P
-Po Pc
c-1K=
o
c
Profe s s i ona l and Re l i ab l e
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Profe s s i ona l and Re l i ab l e
Kappa( K coefficient of agreement; K)
0.81~1 (almost perfect)
0.61~0.80 (substantial)
0.41~0.60 (moderate)
0.21~0.40 (fair)
0.0~0.20 (slight)
K value is between -1 and 1. If the raters are in complete agreement then κ = 1. If there is no agreement among the raters other than what would be expected by chance
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P r o fe s s i o n a l a n d Re l i a b l e
Sentinel ® FMDV NSP Ab ELISA kit
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P r o fe s s i o n a l a n d Re l i a b l e
Sentinel ® FMDV NSP Ab ELISA kit
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Reliability
P r o fe s s i o n a l a n d Re l i a b l e
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Two internal controls (IC-SPC , IC-PC )* were run in 2 replicates on 5 different plates in the same lot.(The data presented here are CV)
within run precision
PI Value Plate-A Plate-B Plate-C Plate-D Plate-E
SPC 0.27% 0.24% 0.00% 0.07% 0.62%
PC 0.31% 0.69% 1.46% 1.31% 2.32%
P r o fe s s i o n a l a n d Re l i a b l e
between run precision Two internal controls (IC-SPC , IC-PC )* were run 10 replicates in the same lot.
*Strong positive control-SPC*positive control-PC
PI Value AVE. STD CV
SPC 96.5 0.31 0.33%
PC 89.0 1.26 1.42%
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different lot precision
Two internal controls (IC-SPC , IC-PC ) were run in PD601631 and PD601648, 2 different lots.
PI Value Ave. STD. CV
SPC 96.4 0.14 0.15%
PC 86.8 3.11 3.58%
P r o f e s s i o n a l a n d R e l i a b l e
When tested the same sample in different lot, the result were high consistency
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P r o fe s s i o n a l a n d Re l i a b l e
Lab tests- Sensitivity and Specificity Animal Health Research Institute (AHRI) provided swine and goat sera than
analyzed.
1. Swine sera30 swine sera infected with O type FMD virus50 FMDV-free swine seraSensitivity: 96.7% (29/30)Specificity: 100% (50/50)
2. Goat sera30 goat sera infected with FMD virus50 FMDV-free goat seraSensitivity: 100% (30/30)Specificity: 100% (50/50)
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Field data
P r o f e s s i o n a l a n d R e l i a b l e
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PrioCHECK®
Total+ -
Sentinel®+ 47 5 52- 4 57 61
Total 51 62 113
113 pigs sera were collected from field farm,Sentinel ® NSP Ab ELISA test kit and PrioCHECK ® NS ELISA were compared
• Observed agreement = 0.92
• Kappa value= 0.83
• Sensitivity※ = 92.20%
• Specificity※ = 91.90%
• ( ※ PrioCHECK ® as standard)
Sentinel ® FMDV NSP Ab ELISA test kit compared with PrioCHECK ® FMDV NS ELISA kit
Profe s s i ona l and Re l i ab l e
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PrioCHECK®
Total+ -
Sentinel®+ 17 2 19- 3 159 162
Total 20 161 181
181 cattle sera were collected from field farm,Sentinel ® NSP Ab ELISA test kit and PrioCHECK ® NS ELISA were compared
• Observed agreement = 0.97• Kappa value= 0.86• Sensitivity※ = 85%• Specificity※ = 98.1%• ( ※PrioCHECK ® as standard)
Sentinel ® FMDV NSP Ab ELISA test kit compared with PrioCHECK ® FMDV NS ELISA kit
P r o fe s s i o n a l a n d Re l i a b l e
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Profe s s i ona l and Re l i ab l e
PrioCHECK®
Total+ -
Sentinel®+ 3 1 4- 1 127 128
Total 4 128 132
132 goats sera were collected from field farm,Sentinel ® NSP Ab ELISA test kit and PrioCHECK ® NS ELISA were compared
• Observed agreement = 0.98
• Kappa value= 0.74
• Sensitivity※ = 75%
• Specificity※ = 99.2%
• ( ※ PrioCHECK ® as standard)
Sentinel ® FMDV NSP Ab ELISA test kit compared with PrioCHECK ® FMDV NS ELISA kit
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Profe s s i ona l and Re l i ab l e
Field tests-validation: The agreement of analytical performance compare to commercial ELISA kit (PrioCHECK)
Species Sample No. Observed agreement Kappa value
Swine 113 0.92 0.83
Cow 181 0.97 0.86
Goat 132 0.98 0.74
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Profe s s i ona l and Re l i ab l e
The performance of our FMDV NSP Ab Testing kits
• Sensitivity• Specificity• Reliability
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Profe s s i ona l and Re l i ab l e
THANK YOU FOR
YOUR ATTENTION
Passion, Integrity, Professional, and EthicsExcelsior Bio-System
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P r o fe s s i o n a l a n d Re l i a b l e