Screening of MG and MS on day old chicken for animal quality control

1. GeneReach
GeneReach
A Fully Automated POCKIT Central PCR
System Detects Mycoplasma
gallisepticum and Mycoplasma
synoviae on DOCs
Simon Chung
GeneReach Biotechnology Corporation

2. GeneReach
Introduction
• Mycoplasma gallisepticum and Mycoplasma synoviae are the main
causative agents of respiratory disease in domesticated birds including
commercial broiler. They are transmitted horizontally and vertically
• Diagnosis of MG and MS by bacteria isolation and serology tests is not
timely for disease treatment in commercial broiler farms
• MG and MS carry over contamination on DOC is suspected as the origin of
outbreak in the farm
• MG and MS are often considered as long lasting pathogens in the
commercial farms and not easy to be cleaned
• Treatment of MG and MS infection only can be processed in the first 3
weeks
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3. GeneReach
Objective
• Prove the utility of POCKITTM Central system on detection of MG and
MS on DOCs tracheal tissue and swab samples
• Try to see if early treatment can be applied to the DOCs contaminated
by MG or MS
• Optimize the detection procedure for DOCs quality control and
disease monitoring
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4. GeneReach
Material and Method of Trial 1
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5 individual tracheal tissue samples are collected from every storey and homogenized by tacoPrep bead beating homogenizer
25,000 DOCs per storey
200 ul of pooled sample is used for PCR testing
100 ul of supernatant is used for sample pooling
Day 0
Day 28
Day 7 Day 14 Day 21

5. GeneReach
Material and Method of Trial 1 (cont.)
Number of PCR Positive result
DOC Day 7 Day 14 Day 21 Day 28
MG MS MG MS MG MS MG MS MG MS
Farm A
(200,000)
Farm B
(200,000)
Farm C
(100,000)
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6. GeneReach
POCKITTM Central system
• Fully automated with easy
sample ID input and data output
• Modulized Nucleic acids extraction
and lyophilized PCR reagents
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Sample-in-Answer-out, just in 85 minutes

7. GeneReach
POCKITTM Central MG and MS Reagents
• Lyophilized format and ready to use
• No cold chain is needed for shipping
• POCKIT MG Reagent targets mgc 2
gene
• POCKIT MS Reagent targets vlhA
gene
• Pre-loaded magnetic beads
DNA/RNA Extraction module
• All plastic consumables are
enclosed in Transfer module
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8. GeneReach
Example of POCKITTM Central Detection Results

9. GeneReach
Results of Trial 1
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Number of PCR Positive result
DOC Day 7 Day 14 Day 21 Day 28
MG MS MG MS MG MS MG MS MG MS
Farm A
(200,000)
0 0 0 0 0 0 0 0 0 0
Farm B
(200,000)
0 0 0 0 0 0 0 3 11 12 No treatment
Farm C
(100,000)
0 0 0 0 0 0 0 0 0 0
Question:
1. Why MS and MG PCR positive results show up since day 21?
2. Did we reduce the detection sensitivity due to the dilution effect of pooling samples?

10. GeneReach
Material and Method of Trial 2
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25,000 DOCs per storey
Day 0
Day 28
Day 7 Day 14 Day 21
Trachea swab
Pool 5 tracheal swabs in
a 15 ml tube containing
3ml taco sample storage
solution
Detect MG and MS
nucleic acids individually
by POCKITTM Central
The results are directly
displayed on the screen
Pool 5 tracheal swabs in a 15 ml tube containing 3ml taco Sample Storage Solution to reduce the dilution effect

11. GeneReach
Material and Method of Trial 2
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Number of PCR Positive result
DOC Day 7 Day 14 Day 21 Day 28
MG MS MG MS MG MS MG MS MG MS
Farm A
(200,000)
Farm B
(200,000)
Farm C
(100,000)

12. GeneReach
Results of Trial 2
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Number of PCR Positive result
DOC Day 7 Day 14 Day 21 Day 28
MG MS MG MS MG MS MG MS MG MS
Farm A
(200,000)
0 0 0 0 1 0 0 1 2 2
Treat with Ampicilin and Colistin
on Day 17
Farm B
(200,000)
1 0 0 0 0 0 1 5 0 2
Treat from Day 2 and stop at
Day14
Farm C
(100,000)
0 0 3 0 0 0 0 0 0 0 No clinical sing no treatment

13. GeneReach
Summary of Trial 1 and Trial 2
• POCKITTM Central System can detect MG and MS from tracheal tissue and
swab samples and deliver the test results at the same day of sampling
• Dilution effect from pooling samples will reduce the detection sensitivity
• In trial 2, MG can be detected from DOCs after revising the sampling
strategy
• Treatment of Farm B in Trial 2 followed by PCR testing tent to delay the
infection of MG
• MS PCR positive results are observed after Day 21 is common in both Trail
1 and 2
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14. GeneReach
Take Home Message
• PCR can be a quality control tool for checking MG and
MS contamination of DOCs
• POCKITTM Central System can be easily adapted by
poultry growers
• Treatment followed by PCR test tent to delay the
outbreak of Mycoplasmas infection
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15. GeneReach
Detecting MG/MS for Cleaning Quality
Control after Harvesting

16. GeneReach
Chang Jhen
Corporation,
Taiwan
Intervet Animal
Health Taiwan
Limited (MSD
Animal Health,
Taiwan), Taiwan
Technical
support
team of
GeneReach
Acknowledgement
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Norman Chen
Leon Lee
Nina Chen
Erin Tsai
Frank Chung

17. GeneReach
GeneReach
Thank You for the Attention
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