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