Rapid and sensitive detection of severe acute respiratory syndrome coronavirus by rolling circle amplification.
The severe acute respiratory syndrome (SARS) epidemic of 2003 was responsible for 774 deaths and caused significant economic damage worldwide. Since July 2003, a number of SARS cases have occurred in China, raising the possibility of future epidemics. We describe here a rapid, sensitive, and highly efficient assay for the detection of SARS coronavirus (SARS-CoV) in cultured material and a small number (n = 7) of clinical samples. Using rolling circle amplification (RCA), we were able to achieve sensitive detection levels of SARS-CoV RNA in both solid and liquid phases. The main advantage of RCA is that it can be performed under isothermal conditions with minimal reagents and avoids the generation of false-positive results, a problem that is frequently encountered in PCR-based assays. Furthermore, the RCA technology provides a faster, more sensitive, and economical option to currently available PCR-based methods.
Wang
Bin
B
Retroviral Genetics Laboratory, Centre for Virus Research, Westmead Millennium Institute, The University of Sydney, Darcy Rd., Westmead, Sydney, NSW 2145, Australia. bin_wang@wmi.usyd.edu.au
Potter
Simon J
SJ
Lin
Yiguang
Y
Cunningham
Anthony L
AL
Dwyer
Dominic E
DE
Su
Yuelong
Y
Ma
Xuejun
X
Hou
Yunde
Y
Saksena
Nitin K
NK
D001483:Base Sequence; D002681:China / Q000453:epidemiology; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis* / Q000401:mortality
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral