Volume 10, Number 7—July 2004
Dispatch
SARS Coronavirus Detection
Table 2
CT values and standard deviation of serial dilutions of SARS-CoV RNA subjected to different real-time RT-PCR assaysa
GE | Real-time RT-PCR systems | Artus kit n = 4 | Drosten et al. (5) n = 6 | NS pp1ab n = 7 | NS pp1abb n = 7 | SS GP b n = 7 | NS pp1a/NS pp1ab/SS GPb n = 6 | |
---|---|---|---|---|---|---|---|---|
500 |
LightCycler |
28.69 ± 0,46 |
28.16 ± 0.04 |
29.57 ± 0.87 |
29.56 ± 0.18 |
29.47 ± 0.53 |
28.53 ± 0.26 |
|
SDS7700 |
n.d. |
27.29 ± 0.09 |
29.63 ± 0.94 |
30.92 ± 0.09 |
26.98 ± 0.65 |
26.11 ± 0.40 |
||
50 |
LightCycler |
32.68 ± 0.26 |
45.00 |
33.85 ± 0.60 |
32.83 ± 0.41 |
32.83 ± 0.41 |
32.48 ± 0.69 |
|
SDS7700 |
n.d. |
31.62 ± 0.45 |
33.82 ± 0.23 |
34.73 ± 1.04 |
30.33 ± 0.26 |
29.29 ± 0.67 |
||
10 |
LightCycler |
34.42 ± 0.20 |
45.00 |
36.72 ± 0.42 |
35.99 ± 0.55 |
35.33 ± 0.48 |
34.50 ± 0.25 |
|
SDS7700 |
n.d. |
38.37 ± 4.02 |
35.69 ± 0.35 |
36.94 ± 0.10 |
32.93 ± 0.81 |
32.01 ± 0.44 |
||
5 |
LightCycler |
35.66 ± 0.40 |
45.00 |
45.00 |
37.61 ± 0.55 |
37.12 ± 0.35 |
37.20 ± 0.30 |
|
SDS7700 | n.d. | 41.15 ± 2.17 | 37.31 ± 0.77 | 37.64 ± 0.67 | 34.55 ± 0.50 | 34.09 ± 0.24 |
aCT, threshold cycle; SARS-CoV, severe acute respiratory syndrome–associated coronavirus; RT-PCR, reverse transcription–polymerase chain reaction; GE, genome equivalents/assay; pp1ab, nonstructural polyprotein 1ab gene; n.d., not determined.
bSee Table 1.
References
- Peiris JS, Lai ST, Poon LL, Guan Y, Yam LY, Lim W, Coronavirus as a possible cause of severe acute respiratory syndrome. Lancet. 2003;361:1319–25. DOIPubMedGoogle Scholar
- Poutanen SM, Low DE, Henry B, Finkelstein S, Rose D, Green K, Identification of severe acute respiratory syndrome in Canada. N Engl J Med. 2003;348:1995–2005. DOIPubMedGoogle Scholar
- Tsang KW, Ho PL, Ooi GC, Yee WK, Wang T, Chan-Yeung M, A cluster of cases of severe acute respiratory syndrome in Hong Kong. N Engl J Med. 2003;348:1977–85. DOIPubMedGoogle Scholar
- Acute respiratory syndrome. China, Hong Kong Special Administrative Region of China, and Viet Nam. Wkly Epidemiol Rec. 2003;78:73–4.PubMedGoogle Scholar
- Drosten C, Günther S, Preiser W, van der Werf S, Brodt HR, Becker S, Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med. 2003;348:1967–76. DOIPubMedGoogle Scholar
- Ksiazek TG, Erdman D, Goldsmith CS, Zaki SR, Peret; Emery S, et al. A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med. 2003;348:1953–66. DOIPubMedGoogle Scholar
- World Health Organization. Case definition for surveillance of severe acute respiratory syndrome SARS. [cited 2003 May 1]. Available from: http://www.who.int/csr/sars/casedefinition/en/
- Poon LL, Wong OK, Chan KH, Luk W, Yuen KY, Peiris JS, Rapid diagnosis of a coronavirus associated with severe acute respiratory syndrome (SARS). Clin Chem. 2003;49:953–5. DOIPubMedGoogle Scholar
- Pratelli A, Martella V, Elia G, Decaro N, Aliberti A, Buonavoglia D, Variation of the sequence in the gene encoding for transmembrane protein M of canine coronavirus (CCV). Mol Cell Probes. 2001;15:229–33. DOIPubMedGoogle Scholar
- Vaughn EM, Halbur PG, Paul PS. Sequence comparison of porcine respiratory coronavirus isolates reveals heterogeneity in the S, 3, and 3-1 genes. J Virol. 1995;69:3176–84.PubMedGoogle Scholar
- Ruan YJ, Wie CL, Ee AL, Vega VB, Thoreau H, Su ST, Comparative full-length genome sequence analysis of 14 SARS coronavirus isolates and common mutations associated with putative origins of infection. Lancet. 2003;361:1779–85. DOIPubMedGoogle Scholar
- Holmes KV, Lai MMC. Coronaviridae: The viruses and their replication. In: Fields BN, Knipe DM, Howley PRM, editors. Fields virology. 3rd ed. New York: Lippincott Williams & Wilkins; 2001. p. 1075–94.
- Chinese SARS Molecular Epidemiology Consortium. Molecular evolution of the SARS coronavirus during the course of the SARS epidemic in China. Science. 2004;303:1666–9. Epub 2004 Jan 29. DOIPubMedGoogle Scholar
- People’s daily edited by ProMED-mail. SARS worldwide. ProMed January 2, 2004. Accessed at: http://www.promedmail.org, archive number: 20040102.0015. 19991013.1826.
- Wood L. questions about comparative genomics of SARS coronavirus isolates. Lancet. 2003;362:578–9. DOIPubMedGoogle Scholar
- Radonic A, Thulke S, Mackay IM, Landt O, Siegert W, Nitsche A. A guideline to reference gene selection for quantitative real-time PCR. Biochem Biophys Res Commun. 2004;313:856–62. DOIPubMedGoogle Scholar
- Kondro W. Canadian officials watch SARS-like mystery bug. Lancet. 2003;362:714. DOIPubMedGoogle Scholar