experience with sars laboratory diagnostics jsm peiris, llm poon, y guan, kh chan. the university of...
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Experience with SARS Laboratory Diagnostics
JSM Peiris, LLM Poon, Y Guan,
KH Chan.
The University of Hong Kong &
Queen Mary Hospital
Early Response: March 28
Ist Generation assays
• RT-PCR for P gene
• Indirect immuno-fluorescence serology using virus infected cells
0 10,000 20,000 29,736 nt
Orf1a polyproteinOrf1ab polyprotein
E2 glycoproteinE1 protein
M proteinN protein
RT-PCR tests: 1st Generation
positive negative H20
2nd Generation RT-PCR testsApril 03
Real Time RT-PCR
Roche Light Cycler
Viral load in Nasopharyngeal aspirates
Peiris et al 2003, Lancet 361: 1767-72
ABI GeneAmp 7000 using TaqMan detection
• Run capacity 96 specimens • Run time 1.5 hrs • Specificty- Probe
• Compare different gene targets• P gene• N gene
10e510e6 10e4 10e3 10e2 10e1 water
TaqMan assay for polymerase gene
R2:0.99
10e6 10e5 10e4 10e3 10e2 10e1